How Our Bodies Adapt to Chemicals - Society of …...1964 • Drugs and other xenobiotics cross membranes due to their lipid solubility. • Drugs are biotransformed to water-soluble

How Our Bodies Adapt to Chemicals

Curtis D Klaassen PhD DABT ATS

bull How Our Bodies Adapt to Chemicals or Developing Resistance to Toxicity or Learning to Reprogram the Liver or Progress Depends on Technology

1964

bull Animal ----------------------------------------------------

bull Analytical methods ------------------------------------ bull Colorimeterspectrophotometer

bull Chromatography bull Paper bull TLC

bull Cell fractions (Centrifuge) --------------------------

bull Null mice and rats bull Transgenic mice

bull Computer ----------------------------------------------- bull Oneuniversity

bull Computer bull More than oneperson

bull Radioactivity bull GC bull HPLC-UV bull HPLC-MSMS

bull Tissue culture

present

bull Antibodies bull Radioimmunoassay bull Western blot

bull mRNA

bull Northern blot bull RT-PCR bull Bead assays bull Microarray bull RNA-Seq

bull Transcription factor ndash DNA binding

bull ChIP-qPCR bull ChIP- Seq

1964 2012

-------------------------------------

-------------------------------------

-------------------------------------

1964

bull Drugs and other xenobiotics cross membranes due to their lipid solubility

bull Drugs are biotransformed to water-soluble compounds so they do not pass cell membranes and thus are excreted more readily

bull Microsomal enzyme inducers increase cytochrome P-450s and enhance the elimination of some drugs

bull There might be two cytochrome p450s

Examples of Liver-Reprogramming

bull P-450 inducers bull Heavy metals bull Plants bull PXR activation bull Development of liver after birth bull Diet

Therefore if the purpose of p450 inducers was to enhance

the elimination of chemicals might the inducers also

increase the elimination of chemicals that are not

biotransformed by enhancing their passage across

membranes

Hypothesis

Plasma clearance of DBSP

Klaassen and Plaa JPET 161361 1968 Time (min)

Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital

LIVER CELL AS THE CENTRAL PROCESSING UNIT FOR DRUGS

AND NUTRIENTS

Bile Blood Programming

Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY

Effect of PCN on Ouabain uptake in isolated rat hepatocytes

Eaton and Klaassen JPET 208381-385 1979

0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



Cd tolerance from Cd pretreatment

Goering and Klaassen JTEH 14803 1984

Rats were pretreated with CdCl2 (20 mg Cdkg sc) and at indicated time

challenged with a lethal dose of CdCl2 (40 mgkg iv) and mortality was

recorded within 48 hrs after Cd challenge

Time after Cd pretreatment n Mortality () No pretreatment 10 100 2 hr 10 100 4 hr 10 100 6 hr 10 60 8 hr 10 50 24 hr 10 0 2 day 10 0 4 day 10 0 8 day 10 0

16 day 10 0

Cd pretreatment protected against acute Cd

hepatotoxicity

Goering and Klaassen TAAP 74308 1984

100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment

Rats were pretreated with CdCl2 (20 mg Cdkg sc) and 24 hrs later were

challenged with hepatotoxic doses of CdCl2 (20-40 mgkg iv) and serum

SDH was determined 10 hrs later Data are Mean plusmn SE of 4-6 rats

Cd pretreatment alters

subcellular Cd distribution


0

10

20

30

40

50

60

70

80

Nuclei Mitochondria Microsome Cytosol

C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment


challenged with hepatotoxic doses of CdCl2 (35 mg Cdkg iv) and subcellular

Cd distribution was determined 2 hrs later Data are Mean plusmn SE of 6 rats

Cd pretreatment alters cytosolic Cd

distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment


challenged with hepatotoxic doses of CdCl2 (35 mg Cdkg iv) and

cytosolic Cd distribution was determined 2 hrs later

MT-null mice are highly susceptible to

chronic Cd-induced nephrotoxicity

LiuhelliphellipKlaassen ToxSci 46 197 1998

Wild-type control and MT-null mice were

given CdCl2 (005-24 mgkg sc) daily for 6

weeks and Blood urea nitrogen (BUN) and

renal MT were determined N =6-8 mice

The relationship between human exposure to

cadmium and renal injury

Klaassen et al Ann Rev Pharmacol Toxicol 39 267 1999



bull In 1987 a post-doctoral fellow Jie Liu came to my Lab with a ldquosuitcaserdquo of chemicals in Traditional Chinese Medicine that he thought would protect against liver injury

Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin

18β-GA 18 β-Glycyrrhetinic acid

18 α-GA 18 α-Glycyrrhetinic acid

HAG 19 α-Hydroxyl asiatic acid 29-0- β-glucoside

HA 19 α-Hydroxyl asiatic acid

Liu and Klaassen Fundam Appl Toxicol 2234-40 1994

CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20

CON Jgs Ngs Ful OA Swe Om DDB


SalineBromobenzeneAcetaminophenCarbon tetThioacetamideFurosemidePhalloidinColchicineCadmiumD-GalNLPS

AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2

Oxidative stress and activators

Nrf2

Anti-oxidant and inflammation Cell survival and proliferation

Drug metabolism Lipid metabolism

Ke

ap1

Ke

ap1

Nrf2

CUL3

Modified from DAutreacuteaux and Toledano Nat Rev Mol Cell Biol 8813 2007

Oleanolic acid is a Nrf2 activator N

rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND

Oleanolic Acid - + - +

Wild-type Nrf2-nullL

iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT

A single dose of oleanolic acid (30mgkg ip) increases Nrf2 translocation into nucleus and induce Nrf2 target genes in wild-type but not Nrf2-null mice

Reisman and Klaassen Biochem Pharmacol 771273-82 2009

What Genes Might the Keap1-Nrf2 Pathway Alter

Nrf2-null Wild-type Keap1-KD Keap1-HKO

Nrf2

Generation of ldquoGene dose-responserdquo Model

Col 2 Col 2 Col 2 Col 2 Nrf2

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA

Wu and Klaassen Toxicol Sci 123590-600 2011

Constitutively Induced or Suppressed Genes

Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6


What Drug Processing Genes are Altered by Nrf2 activation

bull Uptake transporters

bull Phase-I enzymes

bull Phase-II enzymes

bull Efflux transporters

Phase-II enzymes Glutathione Conjugation

Glutathione S-transferase

Gsta2 Gsta3 Gsta4 Gstm1 Gstm2 Gstm3 Gstm4 Gstm6 Gstp1 Gstt3 Mgst3

Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO

Not altered Gstk1 Gstm5 Gstm7 Gsto1 Gstp1 Gstt1 Gstt2

Wu hellip Klaassen PLoS one 7 e39006 2012

Yeager and Klaassen Toxicol Sci 111238 2009

AhR and Nrf2 interactions

UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1

Hydrophilicityeacute

PhaseIIUgtsSults

ElectrophilesOxida veStressand

Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine

γ-glutamylcysteine GSH

Glycine

Gcl Gs Nrf2

Expression of Which Antioxidant Genes are altered by Nrf2

Antioxidant Genes were Induced with Graded Nrf2 Activation

H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr

glutamate cysteine glucine

Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH

Michael Sporn ldquothe father of chemopreventionrdquo asked for our Oleanolic Acid so he could make

more potent analogues

CDDO$ CDDOMe$ CDDOIm$

Oleanolic$acid$



Transcription factor

DNA

RNA

Nucleus

TRANSCRIPTION FACTORS AND GENE REGULATION

Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization

nuclear receptor structurefunction

ligand coactivator

Prototypical Target Genes of Xeno-sensors

Chemicals Transcription Factor

Prototypical target genes in liver

TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11

Clofibrate PPARα Cyp4a14

OA Nrf2 Nqo1

Transporters in Liver B

loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux

Apical SideBasolateral Basolateral

OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4

Klaassen and Lu Toxicol Sci 101 186 2008

Hepatic induction of rat Oatp1a4 protein expression

000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)

AhR CAR PXR PPARα Nrf2

Rausch-Derra helliphellip Klaassen Hepatology 331469 2001

Guo helliphellip Klaassen JPET 300206 2002

Mouse Oatp1a4 induction by PCN is PXR-dependent

0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)

WT mice PXR-null mice

Cheng and Klaassen DMD 341863 2006

Summary of TF-dependent gene battery using wild-type and TF-null mice

Transcription Factor TF-dependent gene battery in liver

AhR Phase I Cyp1a2Cyp2b10Nqo1Aldh1b1 Aldh7a1

Phase II Gstm2 mGst3 Sult5a1 Ugt1a1 Ugt1a6 Ugt1a9 Ugt2b35

Transporters Mrp4 Mate1 Oatp1a1 Oatp1a4

CAR Phase I Cyp1a2 Cyp2b10 Cyp3a11 Nqo1 Aldh1a1 Aldh1a7

Phase II Gsta1 Gsta4 Gstm1 Gstm2 Gstm3 Gstm4 Gstt1 Sult1e1

Sult2a2 Sult3a1 Sult5a1 Papss2 Ugt1a1 Ugt1a9 Ugt2a3 Ugt2b1

Ugt2b34 Ugt2b35 Ugt2b36

Transporters Mrp2 Mrp3 Mrp4 Oatp1a1 Oatp1a4

PXR Phase I Cyp2b10 Cyp3a11 Aldh1a1 Aldh1a7

Phase II Gsta1 Gstm1 Gstm2 Gstm3 Gstm4 Sult2a2 Sult3a1 Ugt1a1

Ugt1a5 Ugt1a9 Papss2

Transporters Mrp2 Mrp3 Oatp1a4 Abcg5

PPARα Phase I Cyp3a11 Cyp4a14 Nqo1 Aldh1a1 Aldh1a7 Aldh3a2 Aldh9a1

Phase II Gsta1 Gstm3 Gstm4

Transporters Mrp4

Nrf2 Phase I Cyp1a2 Cyp2b10 Nqo1 Aldh1a1

Phase II Gsta1 Gsta4 Gstm1 Gstm2 Gstm3 Gstm4 Gstt2 mGst3

Ugt2b35 Ugt2b36

Transporters Mrp3 Mrp4 Oatp1a1

Aleksunes and Klaassen DMD 40 1191 2012

PXR-Response Elements

AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat

Spacers (n=123hellip)

bull It is generally recognized that DR3 and ER6 are the prototypical DNA-binding motifs

for PXR based on data obtained from a few genes

bull Examples CYP3A4 in humans ER6 (AG(GT)TCAnnnnnnAG(GT)TCA) -1kb of TSS

CYP3A23 in rats DR3 (AGTTCAtgaAGTTCA) 5rsquo-flanking region (-133bp of TSS)

El-Sankary et al DMD 28 493 2000

Schuetz et al Mol Pharm 54 1113 1998

Is this still true on a genome-wide scale

Hunting for specific nucleotides targeted by a TF within the entire genome

If typed 1500000 pages

PXR-binding to Cyp3a11 gene locus in mouse liver

Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29

Cui hellip Klaassen Nucleic Acids Res 38 7943 2010

PXR-response element DR3 DR4 etc

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4

Examples for Transporters

PXR-binding to Oatp1a4 gene locus in mouse liver

277

549


PXR-response element DR-9 only Location 10kb upstream of the transcription site

DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)

Novel DR5n+4 Periodic DNA-binding Patterns of PXR


The DR5n+4 Periodic Pattern for PXR-binding to DNA 1 helical turn 34 Aring 10 intervals

Watson and Crick (1953 CSH Symp Quant Biol)

DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4

DNA interacts with PXR in the unit of half a helical turn


The DR5n+4 Periodic Pattern for PXR-binding to DNA

An ldquoAccordionrdquo Model








The binding of TFs to DNA does not explain entirely the re-programming of cells

bull Why not all genes expressed every tissue

bull Not always a good correlation between activation of TFs and function of liver cells

The Freedom of Expression is Under Tight Control by Epigenetic Mechanisms

Epigenetic Factors

bull DNA methylation

bull Histone modifications

bull microRNAs

Functionally relevant modifications to the genome that do not involve a change in the nucleotide sequence

Epigentic events control the freedom of gene expression without altering the underlying DNA sequence

Epigenetics

microRNAs

CpG

DNA methylation Histone modification

RNAPol-II

TFs

RISC

Methylated gene

DNA Methylation as a Suppression Signal for Gene Expression

Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO

Histone Modifications Determine whether the DNA is ldquoLooserdquo Enough to be Accessed by Transcription Factors

Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR

microRNAs usually lead to decreased mRNA stability and suppression of protein synthesis



Comparison of 24hr LD50 of Ouabain in rats of

varying age

( Klaassen JPET 1972)

AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

Differential distribution of Ouabain in Livers of young rats

Three Patterns of Liver Transporters during Development

-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult

CuihellipKlaassenToxicolSci2012

62cri caldrugtransporters



ReprogrammingtheLiverbyDiets

HighCholesterolampCA

HighFat

HighSatFatampSucrose

LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow

5282geneshadalteredmRNAsinatleastonediet

Significance and Conclusion bull Liver can be programmed and re-programmed bull This is the way the liver adapts to its environment bull All billion liver cells have the hardware and software to be re-

programmed bull It is chemicals that alter the programming bull There are no drugs for liver failure bull If we can learn which chemicals will turn on and off the various

programs we will have drugs for liver diseases Cirrhosis and fatty liver Nutritional functions obesity and diabetes Excretory functions jaundice and hyper-bilirubinemia Clotting factors heart attacks stroke Cholesterol synthesis and degradation atherosclerosis

Thank you

curtisklaassenphdgmailcom

bull How Our Bodies Adapt to Chemicals or Developing Resistance to Toxicity or Learning to Reprogram the Liver or Progress Depends on Technology

1964

bull Animal ----------------------------------------------------








bull Tissue culture

present


bull mRNA




1964 2012

-------------------------------------

-------------------------------------

-------------------------------------

1964











membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


1964

bull Animal ----------------------------------------------------








bull Tissue culture

present


bull mRNA




1964 2012

-------------------------------------

-------------------------------------

-------------------------------------

1964











membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



bull mRNA




1964 2012

-------------------------------------

-------------------------------------

-------------------------------------

1964











membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


1964











membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you








membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you






membranes

Hypothesis



Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




Pla

sma

DB

SP C

on

cen

trat

ion

(m

g1

00

ml)

1

10

100

0 10 20 30

Control

Phenobarbital


AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



AND NUTRIENTS


Re-programming

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY



0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




0

05

1

15

2

25

3

35

0 100 200 300 400 500 600 700 800 900 1000

Ouabain (microM)

V 0

Cont PCN

A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


A

BRICK

WALL

PROGRESS

DEPENDS ON

TECHNOLOGY









16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you










16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you








16 day 10 0


hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



hepatotoxicity


100

1000

10000

100000

1000000

Control 20 30 40 50

SDH

(S

un

its

ml)

mg Cdkg

No pretreatment

Cd pretreatment







0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





0

10

20

30

40

50

60

70

80


C

d d

istr

ibu

tio

n in

th

e li

ver

No pretreatment

Cd pretreatment





distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



distribution


0

05

1

15

2

25

3

35

06 10 14 18 22 26

ug

Cd

g li

ver

VeVo

No pretreatment

Cd pretreatment

















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you















Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you








Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Jie Liu 87-98

OA Oleanolic acid

UA Ursolic acid

UV Uvaol

α-H α-Hederin

HD Hederagenin

GL Glycyrrhizin






CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



CCl4

AL

T (

UL

)

0

500

1000

1500

2000

2500

3000

SD

H (

U

ml)

0

5000

10000

15000

20000

25000

NE

CR

OS

IS (

grad

e)

00

05

10

15

20




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




AL

T (

UL

)

0

2000

4000

6000

8000

10000


SD

H (

kIU

L)

0

5000

10000

15000

20000

25000

Saline

Bromo benz ene

Acetaminophen

Carbon tet

Thioacetamide

Furosemide

Phall oidi n

Colchi cine

Cadmium

D-GalNLPS

Ne

cro

sis

(g

rad

e)

0

1

2

3

VehicleOA

Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


Oleanolic acid

ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


ARE

Maf Nrf2


Nrf2



Ke

ap1

Ke

ap1

Nrf2

CUL3



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



rf2 P

rote

in

Rela

tive t

o C

on

tro

l

No

rmalized

to

-a

cti

n

0

1

2

3

4

5

6

7

8

ND ND



iver

mR

NA

Exp

ressio

n(N

orm

alized

to

Co

ntr

ol)

0

2

4

6

8

10

12

14

16

18Ho-1Nqo1 GclcNrf2


nullWT

- + - +

nullWT

- + - +

nullWT

- + - +

nullWT





Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Nrf2



Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Pro

tein

exp

ressio

n

0

1

2

3

4

NA



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Induced genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6

Suppressed genes

Nrf2-n

ull

Wild

-type

Keap1-KD

Keap1-HKO

Fo

ld-i

nd

ucti

on

0

2

4

6










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you










Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





Fo

ld-i

nd

uc

tio

n

0

2

4

6

8

10

30

32

34 Nrf2-null

Wild-type

Keap1-KD

Keap1-HKO





UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


UPTAKEPhaseICyps

Diffusion

BIOTRANSFORMATION

Nucleophiles EFFLUX

Detoxific

aon

PhaseINqo1Eh-1


PhaseIIUgtsSults


Forma onofAdducts

Mrps

GSHConjuga on

Conjugates

Gsts

Glutamate

Cysteine


Glycine

Gcl Gs Nrf2



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


H2O2

H2O

Prdx (red)

Prdx (oxi)

Txn (red)

Txn (oxi)

NADPH

NADP+

Txnrd1

Prdx (red)

Prdx (oxi)

Sulfiredoxin

GSSG

GSH

NADP+

NADPH

Gr


Gcl Gs

Gpx

OH

Fe2+

Ferritin

Biliverdin

Bvrb

Bilirubin

NADP+

NADPH




Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Oleanolic$acid$




DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



DNA

RNA

Nucleus


Transcription

factor

DNA

RNA

Nucleus

DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


DBD LBD

Dimerization


ligand coactivator




TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





TCDD AhR Cyp1a1

PB CAR Cyp2b10

Rifampicin

PCN

PXR Cyp3a11


OA Nrf2 Nqo1


loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



loo

d (

Sin

us

oid

)

Bil

e C

an

ali

cu

lus

Blo

od

(S

inu

so

id)

Mrp4

Abca1

Mrp3

Mrp6

Oatp1a4

Ntcp

Oatp1a1

Oatp1b2

Ent1

Oct1

Oat2

Uptake Efflux


OATP1A2

Mdr2MDR3

Abcg5

Bcrp

Mrp2

Mate1

Mdr1

Bsep

Abcg8Atp8b1

OATP1B1

OATP1B3

Atp7b

Oatp1a4



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



000

010

020

030

040

050

060

070

Oat

p1

a4 p

rote

in (

OD

50m

g p

rote

in)





0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



0

100

200

300

400

CON PCN CON PCN

Oat

p1

a4 m

RN

A e

xpre

ssio

n

(RLU

10

mg

tota

l RN

A)



















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


















Transporters Mrp4



Ugt2b35 Ugt2b36




AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



AGGTCA AGGTCA

TGAACT AGGTCA

AGGTCA TGAACT

Direct repeat

Everted repeat

Inverted repeat












Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Corn Oil

PCN

PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Control

PCN

Cyp3a11

36

31

31

33

35 40

Chr 5

66

34

29

10894

29



PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


PX

R b

ind

ing

fo

ld-e

nri

ch

men

t

Corn Oil

PCN

Oatp1a4



277

549



DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


DR4

DR9

DR14

DR19

DR5n+4

(n=0123hellip)





DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




DR4

DR9

DR14

n=0

n=1

n=2

hellip

(n+1) half turns

DR5n+4
















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you















Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you












Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you









Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you






Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Epigenetic Factors



bull microRNAs



Epigenetics

microRNAs

CpG


RNAPol-II

TFs

RISC

Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


Methylated gene


Un-methylated gene

Gene Activation

Gene Silencing

CH

3

CH

3

CH

3

CH

3

STOP

GO


Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



Me Me

H3K4

Me Me

H3K27

Me

Me Me

H3K27

Me

PcG

Me Me

H3K4

H3K4Me2

H3K27Me3

STOP

GO

microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you


microRNA complex

mRNA

3rsquoUTR





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



varying age


AGE (days old)

3 9 15 21 27 33 39 45

LE

TH

AL

DO

SE

50 (

mg

kg

)

0

20

40

60

80

100

120

140


39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



39 DAY OLD RATS

0 5 10 15 20 25 30 35 40 45

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN

7 DAY OLD RATS

TIME (minutes)

0 5 10 15 20 25 30 35 40 45

TIS

SU

E C

ON

C (

gg

)

0

2

4

6

8

10

12

14

16

18

20

22

KID

NEY

PLASM

A

LUNG

LIVER

MUSC

LE

HEA

RT

SPLEEN

BRAIN



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you



-2 0 1 3 5 10 15 20 25 30 45 60

Age (Days)

Peri

nata

lA

do

les

cen

tA

du

lt

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

20

40

60

80

100

120

Perinatal

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ss

ion

(F

PK

M)

0

200

400

600

800

1000 Adolescent

Age (Days)

-2 0 1 3 5 10 15 20 25 30 45 60

Ge

ne

Ex

pre

ssio

n (

FP

KM

)

0

100

200

300

400Adult







HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you






HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you




HighFat


LabChow

EFADeficient

AIN-93Purified

DHADeficient

HighFructose

DietRestric on

ColorKey

HighLow





Thank you





Thank you


Thank you


Documents

How Our Bodies Adapt to Chemicals - Society of …...1964 • Drugs and other xenobiotics cross membranes due to their lipid solubility. • Drugs are biotransformed to water-soluble