Review Article Autophagy and Liver Ischemia-Reperfusion Injurydownloads.hindawi.com/journals/bmri/2015/417590.pdf · 2019-07-31 · In Vitro and In Vivo Animal Studies. e mammalian

Review ArticleAutophagy and Liver Ischemia-Reperfusion Injury

Raffaele Cursio12 Pascal Colosetti2 and Jean Gugenheim1

1Service de Chirurgie Digestive et Transplantation Hepatique Hopital lrsquoArchet 2 Universite de Nice Sophia Antipolis151 route Saint Antoine de Ginestiere 06200 Nice Cedex 3 France2Inserm U1065-C3M Equipe 2 Universite de Nice Sophia Antipolis 06204 Nice Cedex 3 France

Correspondence should be addressed to Raffaele Cursio cursiounicefr

Received 13 June 2014 Revised 21 August 2014 Accepted 7 September 2014

Academic Editor Han-Ming Shen

Copyright copy 2015 Raffaele Cursio et alThis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Liver ischemia-reperfusion (I-R) injury occurs during liver resection liver transplantation and hemorrhagic shockThemainmodeof liver cell death after warm andor cold liver I-R is necrosis but other modes of cell death as apoptosis and autophagy are alsoinvolved Autophagy is an intracellular self-digesting pathway responsible for removal of long-lived proteins damaged organellesand malformed proteins during biosynthesis by lysosomes Autophagy is found in normal and diseased liver Although dependingon the type of ischemia warm andor cold the dynamic process of liver I-R results mainly in adenosine triphosphate depletionand in production of reactive oxygen species (ROS) leads to both a local ischemic insult and an acute inflammatory-mediatedreperfusion injury and results finally in cell death This process can induce liver dysfunction and can increase patient morbidityand mortality after liver surgery and hemorrhagic shock Whether autophagy protects from or promotes liver injury followingwarm andor cold I-R remains to be elucidated The present review aims to summarize the current knowledge in liver I-R injuryfocusing on both the beneficial and the detrimental effects of liver autophagy following warm andor cold liver I-R

1 Introduction

Partial or complete interruption of the liver blood flow andconsequently interruption of its oxygen supply followed byreperfusion and reestablishment of blood flow and oxygensupply characterizes the liver ischemia-reperfusion (I-R)process The cellular injuries caused by the ischemic periodare aggravated by reperfusion [1ndash4] Not only liver transplan-tation or resection but also liver injury following blunt orpenetrating abdominal trauma [5] and hemorrhagic shock[6] may cause low liver blood flow resulting in insufficientperfusion Finally after reperfusion liver I-R injury occursLiver I-R injury following hemorrhagic shock remains amajor cause ofmorbidity andmortality after trauma [6] LiverI-R rapidly leads to an acute inflammatory response causingsignificant hepatocellular damage and organ dysfunctionThe severity of injury ranges from moderately raised levelsof serum aminotransferases to posthepatectomy insufficiencyafter liver resection or to primary nonfunction or initialpoor graft function after liver transplantation [7ndash13] Underextreme circumstances multiple organ failure and deathmay

occur More precisely in liver transplantation up to 10 ofearly transplant failures are caused by I-R injury with a higherincidence of both acute and chronic graft rejection increasingthe need for retransplantation [9ndash13] Liver I-R may also beresponsible for ischemic-type biliary lesions (ITBls) [14ndash16]and late allograft failure [17 18] The use of marginal livergrafts from non-heart-beating donors older andor steatoticorgans and of organs that have been subjected to prolongedperiods of warm ischemia and cold storage has increased inthe last years due to organ shortage [19]These grafts aremorevulnerable to warmcold I-R injury [19] underlining the needof therapeutic strategies to reduce liver I-R injury in order toimprove graft viability [20]

Necrosis represents the main mode of liver cell deathfollowing warmcold I-R [21ndash23] but other modes of celldeath namely apoptosis [2 24 25] and autophagy [26ndash48]also play an important role

Mammalian autophagy is an intracellular self-digestingpathway responsible for the removal of long-lived proteinsdamaged organelles and malformed proteins during biosyn-thesis by lysosomes [49] Autophagy is divided into three

Hindawi Publishing CorporationBioMed Research InternationalVolume 2015 Article ID 417590 16 pageshttpdxdoiorg1011552015417590

2 BioMed Research International

main types according to the different pathway by which cargois delivered to the lysosome or vacuole chaperone-mediatedautophagy (CMA) microautophagy and macroautophagy[50]The last type can be divided into six principal steps initi-ation nucleation elongation closure maturation and degra-dation or extrusion [49] Through these steps cytoplasmicmaterials such as protein aggregates and organelles aresequestered by the phagophore a preautophagosomal mem-brane structure which thereafter expands and encloses itscargo to form a double-membrane vesicle the autophago-some [51] By its fusion with a lysosome it forms an autoly-sosome in which the enclosed cargo is degraded by acidhydrolases into biologically active monomers such as aminoacids that are subsequently reused to maintain cellular meta-bolic turnover and homeostasis [51]

Autophagy is involved in normal and diseased liver [52ndash55] Studies on autophagy in liver tissue following warmcold I-R remain controversial Some show defective andordecreased autophagy [27ndash30 32 34ndash37 39ndash41 45 47] in oth-ers hepatocellular autophagy is increased [26 31 33 38 42ndash44 46 48] Whether autophagy protects from or promotesliver injury following warm andor cold I-R remains to beelucidated

2 Pathomechanisms of Warm andor ColdLiver I-R Injury

Warm ischemia of the liver is due to oxygen deficiency causedby vascular occlusion of the liver during liver resection orhemorrhagic shock [2 10] Cold ischemia is observed duringliver transplantation in which the graft is subjected to warmand cold ischemia followed by a warm reperfusion phase thewarmcold ischemia sequence is due to vascular occlusion ofthe liver graft during its procurement from the donor andto graft storage in cold preservation solutions before livertransplantation [2 10] Graft implantation into the recipientrepresents the warm reperfusion phase all these phases leadto I-R injury of the liver [2 10] Pathomechanisms associatedwith warm I-R seem to differ from those after cold I-R in livertransplantation [57] Unlike warm liver I-R which resultsin primary damage to the hepatocytes cold I-R is mainlycharacterized by injury to the sinusoidal lining cells [58ndash60]

The introduction of oxygenated blood during reperfusionaggravates the ischemic insult which itself is mainly charac-terized by cellular adenosine triphosphate (ATP) depletionand results in perturbation of the cellular energy-dependentmetabolic and transport processes [1 61] Although the graftmetabolism is reduced during hypothermia with a prolongedtime period in which the anoxic cells can retain essentialmetabolic functions hypothermia may induce liver injury bydysfunction of the NaK ATPase membrane pump [61 62]The resulting intracellular influx and accumulation of sodiumand chloride lead to a perturbation of calcium homeostasisand to cell swelling [61 62] The inflammatory response toliver I-R involves neutrophils cytokines chemokines com-plement monocytes and macrophages [63ndash67] The reper-fusion process consists of two phases in the initial phaseactivated resident macrophages of the liver the Kupffer cells

induce oxidative stress mainly by reactive oxygen species(ROS) generation and in the later phase 6ndash24 hours follow-ing reperfusion recruited neutrophils release inflammatorymediators which can cause direct tissue damage [1] Kupffercells play a central role in the pathophysiologicalmechanismsof liver I-R Activated Kupffer cells release both ROS andcytokines including tumor necrosis factor alpha (TNF120572)Interleukin-1 (IL-1) and Interleukin-6 (IL-6) leading to gran-ulocytes accumulation in the sinusoidal space and causingmicrocirculatory disturbances [68] Accumulation of acti-vated neutrophils through release of oxidants and proteasesleads to hepatocyte damage ROS stimulate endothelial cellsto secrete platelet activating factor (PAF) [69] Monocytesand Kupffer cells-derived ROS activate redox-sensitive tran-scription factors AP-1 and nuclear factor kappa-B (NF-120581B) inendothelial cells and hepatocytes [65] Complement activa-tion products activate Kupffer cells during the initial phaseof liver injury and contribute to tissue inflammation as amembrane-attacking complex that stimulates the productionof proinflammatory cytokines and chemotactic agents whichoccur immediately after reperfusion [63] Complement canalso regulate the adaptative immunity [70 71] In fact theinflammation occurring during liver reperfusion is predomi-nantly an innate-immune-dominated response which mightinduce I-R injury in both parenchymal and nonparenchymalcells in situ and in liver transplants [66 67 72 73] Howeverthe cold preservation injury of the graft causes a likewisestrong adaptative immune response characterized by an earlyand massive T-cell influx into the ischemic liver graft [73ndash75]The conventional T lymphocytes CD4+ cells accumulatein the liver within 1 hour after reperfusion preceding anyneutrophil accumulation [76] CD4+ cells are activated byvarious Kupffer cell-derived products and lead to hepatocytesand sinusoidal endothelial cells damage [76] and finally livercells necrosis [22 23 77] Apoptosis seems to be a relevantdeath mechanism during warmcold liver I-R injury too[2 24 25] even if the studies are controversial on this issue[21 24 25] Recently the role of liver autophagy followingwarm andor cold liver I-R has been highlighted [78]

3 Autophagy and Warm andor ColdLiver I-R Injury

Beside necrosis [21ndash23] there are other modes of cell deathas apoptosis [2 24 25] and autophagy that may occur simul-taneously and or sequentially [26ndash48 78] following warmandor cold liver I-RWhile some studies have shown a defec-tive autophagy in hepatocytes following anoxiareoxygen-ation and in liver tissue following I-R [27ndash30 32 34ndash37 39ndash41 45 47] in others an increase of autophagy has beenobserved [26 31 33 38 42ndash44 46 48] (Table 1) with differenteffects on warm andor cold liver I-R injury (Figure 1)

31 Protective Role of Autophagy against Warm andorCold Liver I-R Injury

311 In Vitro and In Vivo Animal Studies The mammalianorthologue of yeast autophagy-related gene 6 (Atg6) Beclin 1

BioMed Research International 3

Table1Au

toph

agyandwarm

andor

cold

liver

I-Rinjuryinvitro

andin

vivo

anim

alandhu

man

studies

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Partialw

arm

60min

Decreased

(a)H

emin

+Ch

loroqu

ine

(b)H

emin

+Wortm

annin

Detrim

ental

Detrim

ental

Yunetal

2014

[47]

Mice

No

Partialw

arm

60min

Increased

Melaton

inProtectiv

eKa

ngetal

2014

[48]

Mice

No

Partialw

arm

45min

Increased

Ethylpyruvate

Protectiv

eShen

etal

2013

[46]

Mice

No

Totalw

arm

45min

Decreased

Carbam

azepine

Protectiv

eKim

etal

2013

[45]

Rat

No

Partialw

arm

60and

90min

Increased

Chloroqu

ine

Protectiv

e(earlyreperfusionph

ase)

Detrim

ental

(latereperfusionph

ase)

Fang

etal

2013

[44]

Rat

No

Partialw

arm

90min

Increased

Chloroqu

ine

Chloroqu

ine+

NAC

Detrim

ental

Protectiv

eSunetal

2013

[43]

Rat

No

Partialw

arm

60min

Increased

Lithium

Protectiv

eLiuetal

2013

[42]

Rat

Steatosis

Coldisc

hemia24

hDecreased

Mela

tonin+Trim

etazidine

Protectiv

eZa

oualietal

2013

[41]

Rat

Steatosis

Coldisc

hemia16h30

min

warm

reperfusionin

situ

Decreased

Detrim

ental

Gracia-Sancho

etal2013[40]

Hum

anNo

Hypoxiareoxygenatio

nof

hepatocytes

Decreased

3-MA

Protectiv

eBh

ogaletal

2012

[39]

Mice

No

(a)A

noxiareoxygenation

ofhepatocytes

Increased

Rapamycin

Protectiv

eWangetal

2012

[38]

(b)P

artia

lwarm

90min

Increased

Rapamycin

Protectiv

eCa

lcium

Calm

odulin-

kinase

IVKO

mice

No

Partialw

arm

60min

Decreased

Rapamycin

Protectiv

eEv

ankovich

etal2012[37]

Hum

anNo

Livertransplantatio

nDecreased

Ischem

icprecon

ditio

ning

Protectiv

eDegliEspo

stietal2011[35]

Old

mice

No

Totalw

arm

20min

Decreased

(a)A

LLM

Calpain2inhibitor

(b)A

deno

viraloverexpression

ofAtg4BandBe

clin1

Protectiv

eWangetal

2011[34]

Pig

No

Livertransplantatio

nDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2011[36]

Rat

Steatosis

Coldisc

hemia20

hDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2009

[30]

Mice

No

Partialw

arm

60min

Decreased

Cisplatin

Protectiv

eCardinaletal

2009

[32]


Table1Con

tinued

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Livertransplantatio

nIncreased

(a)W

ortm

annin

(b)L

Y294002

Protectiv

eGotoh

etal

2009

[33]

Hum

anPo

stchemotherapy

steatosis

TotalW

arm

Decreased

Ischem

icprecon

ditio

ning

Protectiv

eDom

artetal

2009

[29]

Old

mice

No

Partialw

arm

3060and

90min

Decreased

Rosig

litazon

eProtectiv

eSh

inetal

2008

[28]

Rat

No

(a)T

otalwarm

45min

Decreased

Protectiv

eKim

etal

2008

[27]

(b)A

noxiareoxygenation

ofhepatocytes

Decreased

(a)N

utrie

ntdepletion

(b)A

deno

viraloverexpression

ofAtg7

andBe

clin1

(c)A

LLM

Calpain

2inhibitor

Rat

No

Partialw

arm

120m

inIncreased

Protectiv

eCu

rsio

etal

2010

[31]

Rat

No

Livertransplantatio

nIncreased

Detrim

ental

Luetal

2005

[26]


Warm andor cold liver I-R injury

Decreased autophagy

Increased autophagy

Beneficial effects of increased autophagy

- Misfolded protein clearance and removal of damaged organelles

- Recycling of catabolites to maintain cellular energy state by ATP production

Detrimental effects- Excessive degradation of essential

proteins and organelles- Cross-talk to other forms of cell death

apoptosis and necrosis

Kupffer cells activation

Complement activation

CD4 T-cell recruitment and

activation

PI3K classes I and III inhibitors

- 3-MA- Wortmannin- LY294002

Autophagosome-lysosomeinhibitor

Improvement of liver I-R injury

Surgical techniques- Hypothermic reconditioning- Ischemic preconditioning Chemical inhibitors

- Ethyl pyruvate- NAC

Chemical stimulators- Rosiglitazone - Trimetazidine- Simvastatine- Cisplatin

- Nutrient depletion - Short fasting

IMPase inhibitors- Lithium chloride- Carbamazepine

mTOR inhibitor- Rapamycin

Adenoviral overexpressionof Atg7 or Beclin 1

ALLM Calpain 2 inhibitor

ROS TNF120572IL-1120573 and IL-6

uarr MPTuarr ER stress

darr ATP

darr Atg7darr Beclin 1

darr AMPKuarr mTOR

uarr HMGB1

- Chloroquine

Figure 1 Pathomechanisms of warm andor cold liver I-R injury and effects of modulation of autophagy Pharmacological andor surgicalmodulation of decreased or excessive autophagy during warm andor cold liver I-R may improve liver injury Arrow stimulation horizontalT inhibition

has an important role in autophagosome formation as a com-ponent of multiprotein class III phosphatidyl inositol-3 kin-ase (PI3K) complex [79] Beclin 1 is important for localizationof autophagic proteins to preautophagosomal membranestructure during the nucleation step of autophagy [80]Reduced Beclin 1 levels have been observed in hepatocytesduring anoxiareoxygenation in mice [37] and following45min warm liver I-R in rats [27] Increased expression ofthe autophagic protein Beclin 1 by nutrient deprivation by itspharmacologic induction and by its adenoviral overexpres-sion during anoxiareoxygenation in mouse hepatocytes invitro [37] and in rat livers following I-R injury in vivo [32]

protects hepatocytes from cell death and reduces liver I-Rinjury [27 32 37]

Impaired liver autophagy seems to be mediated by cal-cium overload and consequent Calpain 1 and Calpain 2 acti-vation which mediates the proteolytic clivage of the autoph-agic protein Beclin 1 andor of Atg7 in anoxic hepatocytes[27] and in livers following total warm I-R [27 45] Defec-tive autophagy may culminate in onset of mitochondrialpermeability transition (MPT) and hepatocyte death afterreoxygenation [27] MPT results in either necrosis by uncou-pling of oxidative phosphorylation and apoptosis by releasingproapoptotic factors that are normally sequestered in the


mitochondrial intermembrane space [77 81] MPT acts alsoas a molecular signal initiating the autophagic degradation ofmitochondria the mitophagy [82] Impaired mitophagy fol-lowing liver I-R fails to remove dysfunctional mitochondriathe mitochondria loaded with ROS undergo MPT whichin turn leads to uncoupling of oxidative phosphorylationenergetic failure ATP depletion and ultimately cell death[27 82] During liver I-R Kupffer cells neutrophils andplatelets are activatedTheir activation results in a generationand release of ROS and in a cascade of inflammatoryevents including release of proinflammatory cytokines suchas TNF120572 IL-2 IL-6 IL-1 and high mobility group box 1(HMGB1) protein [83] HMGB1 is a DNA binding proteinwhich when secreted actively by nonparenchymal liver cells(Kupffer cells and endothelial cells) and by neutrophils orwhen passively released by necrotic liver cells [84 85] mayinduce an inflammatory signaling cascade [86] HMGB1acts as an alarmin an alarm protein signal that initiatesthe inflammatory response resulting from liver I-R [87] Innormal rat liver HMGB1 is mainly present in the nucleiof hepatocytes [87] HMGB1 was released into the effluentcollected from the infrahepatic vena cava during prolongedcold saline preservation of isolated rat liver grafts [88] Aftercold liver graft preservation for 6 hours and transplantationin rats serum levels of HMGB1 were increased in the earlyreperfusion phase [89] Following 60warmpartial liver I-R inmice [86] and following 90min warm partial liver I-R in rats[90] HMGB1 translocated from the nucleus to the cytoplasmof hepatocytes and was released into the blood circulationwithin 1 hour after reperfusion During warm liver I-R tissuelevels of HMGB1 increase with its innate immune activationrequiring toll-like receptor 4- (TLR4-) dependent signalingalready 1 hour following reperfusion and continue to increasefor up to 24 h later [87] In human liver transplantationthe peak of HMGB1 serum levels was observed 10min afterreperfusion thereafter it started to decrease progressivelywithin 1-2 hours [91] In vitro nontoxic concentrationsof Cisplatin a platinating chemotherapeutic can sequesterHMGB1 inside the nucleus of hypoxic rat hepatocytes canincrease Beclin 1 expression can modulate liver I-R-inducedMAPK activation and can induce autophagy [32] Theseabilities of Cisplatin had beneficial effects on warm liver I-Rinjury and were also observed in vivo in mice [32] In fact invivo administration of nontoxic concentrations of Cisplatinprevented HMGB1 release induced by 60min partial warmliver I-R and reduced subsequent liver injury in mice [32]Liver I-R alone increased Beclin 1 and Atg8microtubule-associated protein 1 light chain 3 (LC3) expression LC3 is alsoinvolved in the formation and expansion of the autophago-some however after Cisplatin administration this increasewas more pronounced and associated to mitophagy andfinally proved to be protective against liver I-R injury [32]

The processing and degradation of LC3 from the uncon-jugated form microtubule-associated protein 1 light chain1 (LC3-I) to the conjugated form microtubule-associatedprotein 1 light chain 2 (LC3-II) which remains associatedto the autophagolysosome indicate an increase in autophagy[49ndash51]

In rats 60min partial warm liver ischemia resulted inincreased liver LC3-II LC3-II is associated with the autoph-agosomal membrane allowing the closure of the autophagicvacuole and increases Atg5 expression which when associ-ated in a protein complex with Atg12 and Atg16 leads toautophagosome formation and finally to increased autophagyduring reperfusion [42] This liver I-R-induced LC3-II andAtg5 expression was more pronounced after chronic Lithiumpretreatment of rats [42] In fact Lithium can induceautophagy by inhibiting inositol monophosphatase (IMPase)and leads to free inositol depletion which in turn maydecrease myo-inositol-145-triphosphate (IP3) levels [92]Induction of liver autophagy by chronic Lithium treatmentbefore induction of 60min partial warm ischemia was asso-ciated with reduced I-R liver injury lower hepatic inflam-matory cytokines levels less liver neutrophil infiltration andlower liver HMGB1 expression and serum HMGB1 levels[42]

It has been shown that the serinethreonine kinase Aktalso known as protein kinase B (PKB) plays a key role incell survival and proliferation and may protect against liverI-R injury [93ndash96] Hepatocytic anoxiareoxygenation and90min partial warm liver I-R in mice resulted in a moderatebut significant increase of hepatocellular levels of autophagy[38] Hydrogen sulphide (H

2S) pretreatment of mice exerted

a protective effect in both hepatocytic anoxiareoxygenationand 90min partial warm liver I-R injuries through Akt1activation [38] However H

2S pretreatment suppressed the

moderate but significant increase of liver autophagy levelsoccurring during hepatocyte reoxygenation and duringwarm liver reperfusion In both experimental conditionspretreatment with the autophagic inducer Rapamycin whichinduces autophagy by binding with cochaperone immuno-philin FKBP1 to specifically inhibit the mammalian target ofRapamycin (mTOR) complex 1 can reverse the autophagicinhibition of H

2S and enhances its hepatoprotective effects

[38]Reducing ROS-induced hepatocellular necrosis seems to

be another protective role of autophagy in 90min partialwarm liver ischemia injury in rats [43] Following 90min par-tial warm liver ischemia without reperfusion both increasedlevels of LC3-II and increased number of autophagosomeswere observed [43] These increases were associated withgeneration ofmitochondrial ROS and liver injury [43]Whenautophagy was inhibited by the autophagy inhibitor Chloro-quine the increase of mitochondrial oxidative stress of ROSproduction and of mitochondrial damage was even morepronounced [43] The consequent accumulation of damagedmitochondria which are normally sequestered and degradedthrough autophagy leads to an enhanced ROS productionwith a subsequent acceleration of ischemia-induced liverinjury and an increase of hepatocellular necrosis [97]

Decreased autophagy is a physiological consequence ofaging [98] In vitro hepatocyte I-R and in vivo warm liverI-R injury associated with autophagy seems to be age-dependent [28 34] In vitro old mice hepatocytes subjectedto 2 hours of hypoxia followed by 12 hours of reoxygenationand in vivo 20min total warm liver ischemia followed by40min reperfusion showed an increase in Calpain 2 activity


which hydrolyzed Atg4B and led to impaired autophagosomeformation impaired autophagic flux and mitophagy andpromoted the onset of the MPT and subsequent cell death[34] The activation of peroxisome proliferator-activatedreceptor gamma (PPAR120574) which belongs to the hormonenuclear receptor superfamily is downregulated during liverischemia aggravating liver injury [99] PPAR120574 activation waslost in old mice after 30min partial warm liver ischemia [28]This suppression of PPAR120574 activation was accompanied by areduced liver autophagy which was more important in cor-relation with the duration of ischemia Pretreatment of micewith the autophagy inducer Rosiglitazone activated PPRA120574and increased liver autophagy after warm ischemia [28]

Steatotic livers have an increased risk of postoperativecomplications after liver resection [100] and liver transplanta-tion [101] They are particularly susceptible to mitochondrialalterations after storage in cold preservation solutions fortransplantation [102 103] Steatosis may provide a substratethat promotes not only oxidative stress but indirectly alsooxidant injury by decreasing the autophagic function [104105] An impaired autophagy was observed during cold I-R of steatotic rat livers [40 41] Pretreatment with someautophagic stimulators as well as Trimetazidine and Simvas-tatine increased liver autophagy and improved I-R injuries ofsteatotic rat livers submitted to cold I-R [40 41]

The use of hypothermic reconditioning (HR) a surgicaltechnique consisting of temporary hypothermic oxygenationof the grafts by using an oxygenated machine perfusion orgaseous oxygen persufflation may improve graft functionand viability [106 107] HR of steatotic livers by insufflation ofgaseous oxygen via the cava vein during the last 90minutes ofcold preservation of the graft has limited mitochondrial dys-function and restored basal rates of hepatocellular autophagyin rats [30] The beneficial effects of HR on liver grafts havealso been shown in transplantation of nonsteatotic pig liverswhere hepatocellular autophagy was preserved mitigatingthe activation of innate immunity and leading to an improvedsurvival of recipients [36]

312 InVitro and InVivoHuman Studies In primary isolatedhuman hepatocytes autophagy is a cell survival mecha-nism during oxidative stress [39] Isolated primary humanhepatocytes which were exposed ex vivo to hypoxia andhypoxia-reoxygenation showed an increase of autophagywithin the mitochondria [39] Inhibition of autophagy by 3-methyladenine (3-MA) in these stressed hepatocytes resultedin the lowering of MPT and onset of cell death by apoptosis[39] During warm and cold liver I-R ROS are responsiblefor lipid peroxidation protease activation cytokine releaseadhesion molecule expression microcirculatory failure andfinally apoptosis and necrosis [63] However ROS are alsocritical mediators of autophagy [108] and during hypoxia-reoxygenation of primary human hepatocytes inhibition oftheir production by N-acetylcysteine (NAC) Rotenone andDiphenyliodonium suppressed autophagy and led to reducedlevels of apoptosis and necrosis [39] Thus ROS seem tobe key mediators of autophagy during oxidative stress anddepending on either the absolute level of intracellular ROSthe type of ROS subspecies generated or the duration of ROS

generation they may be critical factors in determining celldeath by apoptosis or necrosis [22 23 39]

Ischemic preconditioning (IP) of the liver is a surgicalprocedure consisting of a short period of liver ischemia (10min) followed by reperfusion (10 min) Then the prolongedperiod of ischemia by clamping the hepatic artery and theportal vein on the hepatic pedicule (Pringle maneuver) isbetter supported by the liver [109] After promising resultsin animal models IP was efficiently used in clinical studies[110 111] However its benefit to protect the liver from I-R injury in liver resection and transplantation in humansremains controversial [112ndash115] A recent meta-analysis ofIP for liver resection in patients with and without chronicliver diseases failed to find a significant benefit of IP in liverresection [115] Also in liver transplantation there were noclear benefits of IP [112 113] Hepatosteatosis and vascularinjury induced by chemotherapy can reduce tolerance of theliver to reperfusion injury and increase the risk of subsequentliver failure [116 117] In steatotic human livers formerlytreated by chemotherapy the use of IP before prolongedischemia required by liver resection resulted in limitedhepatocyte necrosis and was associated with an activation ofliver autophagy [29] The beneficial effects of IP on liver I-Rinjury seem to be a consequence of autophagy onset leadingto preserved ATP levels and avoiding hepatocellular necrosisby delaying proapoptotic effects [29] In liver transplantationIP of steatotic grafts (preconditioning in the donor beforegraft removal) induced autophagy limited necrosis in humanrecipients and decreased the incidence of rejection episodes[35] In the ischemic preconditioned steatotic graft a cellularincrease of Beclin 1 and LC3 was observed compared withnon-IP steatotic liver grafts [35] In addition there was aninverse correlation between the number of LC3-positive cellsand the necrotic index in IP steatotic liver grafts [35] IPdecreased the incidence of both acute and chronic liver rejec-tion in recipients of steatotic grafts compared to recipients ofnon-IP steatotic grafts [35]

Overall these studies showed that restoration or enhance-ment of autophagymay improve liver I-R injury by providingcells with the energy derived from lysosomal degradation ofcellular materials [118]

32 Detrimental Role of Autophagy inWarmandor Cold LiverI-R Injury

321 In Vitro and In Vivo Animal Studies Several studieshave shown increased levels of autophagy in hepatocytes fol-lowing anoxiareoxygenation and in livers following warm orcoldwarm I-R [26 31 33 38 42ndash44 46 48]These increasedlevels of autophagy appeared either detrimental [26] prob-ably protective [31] or both protective in the early phase ofreperfusion and detrimental in the late phase of liver reperfu-sion [44] Furthermore when pharmacologically modulatedinhibited [33 46 48] or stimulated [38 42 43] autophagyimproved warm andor cold liver I-R injury in both cases

Increased autophagy was observed following partial120min warm liver ischemia in rats 6 hours after reperfusion[31] The hepatocytes of the ischemic liver lobes 6 hours


after reperfusion had dense bodies and various autophago-somes as well as oval and rounded mitochondria [31] Thenumber of hepatocytes with punctate LC3 staining in thecytoplasm was markedly increased in ischemic comparedto nonischemic liver lobes [31] During orthotopic LT inrat autophagosomesautolysosomes were observed in grafthepatocytes in both cold preservation and reperfusion phases[26] Induction of autophagy was more pronounced ingraft hepatocytes after 30 to 60min of warm reperfusionthan in hepatocytes after cold preservation [26] Abundantautophagosomesautolysosomes were associated with dyinghepatocytes within 2 hours of warm reperfusion [26] Warmreperfusion phase may facilitate autophagosome formationin hepatocytes under ATP exhaustion as a stress response[26] About 15 minutes after the start of warm reperfusionsmall masses of hepatocytes with abundant autophagoso-mesautolysosomes frequently dissociated from the hepaticcords and were extruded into the sinusoidal lumen [33]Occlusion of the sinusoidal stream contributed to a mas-sive necrosis of hepatocytes within 2 hours and led toliver dysfunction [33]The hepatocytes containing numerousvacuolarlysosomal structures often underwent degenerationandwere phagocytosed byKupffer cells late in the reperfusionphase [26] The inhibition of autophagosome formation andmaturation by adding the autophagic inhibitor Wortman-nin and LY294002 a specific inhibitor of PI3KAkt kinasepathway to the cold preservation solution attenuated liverdysfunction and recipient mortality rates [33]

The beneficial effects of Rapamycin by reversing theautophagic inhibition of H

2S during hepatocytic anoxia

reoxygenation and 90min partial warm liver I-R inmice [38]are in contrast with the detrimental effects of Rapamycinon reperfused livers following partial warm 60min liverischemia in mice shown in a recent study [48] In fact theincreased levels of autophagy induced by warm reperfusionwere even higher after Rapamycin pretreatment and thisexcessive activation of autophagy aggravated liver I-R injury[48] Furthermore in the same study Rapamycin reversedthe beneficial effects ofMelatonin administration a lipophilicindole secreted by pineal and nonpineal cells which seems toprotect against liver I-R injury by inhibiting oxidative stressand by improving both mitochondrial respiration and ATPsynthesis after cold storage of the liver [119 120] In this studyMelatonin downregulated autophagy via activation of mTORsignaling and resulted in improvement of liver I-R injury [48]

In contrast in moderate and advanced steatotic cold-stored and warm-reperfused livers in rats in which auto-phagy was impaired [40] Melatonin associated with Trime-tazidine induced liver autophagy and improved liver injury[41] When adding Simvastatine a statin possessing vasopro-tective properties to the cold-storage solution the bioavail-ability of the vasoprotector NO was maintained and led toautophagy induction [40] Simvastatin treatment preventedhepatic endothelial dysfunction not only in steatotic [40] butalso in nonsteatotic [121] livers and resulted in improvementof liver injury

An increased level of autophagy was observed also inrat liver following 60 and 90min partial warm liver I-R[44] LC3-II protein was increased at 6 hours after liver

reperfusion this increase was more pronounced when ratswere pretreated with the antimalaria drug Chloroquine [44]Chloroquine seems to have a dual role in warm liver I-R infact it improved liver injury in the early phase of reperfusionby reducing inflammatory cytokines as well as IL-6 IL-1and TNF120572 it diminished HMGB1 release and it modifiedI-R-induced MAP kinase activation [44] In the late phaseof reperfusion however Chloroquine inhibited autophagyand induced apoptosis aggravating liver I-R injuries [44] Bycontrast in another study levels of autophagy were decreasedearlier in hepatocytic anoxiareoxygenation and at 1 and 4hours following partial warm 60min liver I-R in vivo in rats[47] and after Chloroquine treatment liver I-R injuries wereaggravated at these time points studied [47]

HMGB1 has also an important functional role in cross-regulating apoptosis and autophagy [80] Actively secreted bynonparenchymal liver cells Kupffer and sinusoidal endothe-lial cells andor passively released by necrotic liver cells[84 85] the stress sensor with redox activity HMGB1 actsas protein signal that initiates the inflammatory responseresulting from liver I-R [87] For HMGB1 translocation fromnuclei to cytoplasm and for enhanced autophagy ROS signalsare required [122] Following 45min warm partial liver I-R injury in mice HMGB1 translocated from the nucleusto the cytoplasm of hepatocytes competitively combinedwith Beclin 1 and promoted the levels of autophagy throughrepresenting the site of the antiapoptotic Bcl-2 protein whichnormally maintains the inactive status of autophagy andcaused warm liver I-R injury in rat [46]

Ethyl-Pyruvate a lipophilic ester derived from the endog-enous metabolite pyruvate seems to improve liver injury byinhibiting the intrinsic pathway of apoptosis and autophagy[46] Ethyl-Pyruvate might decrease separately both apop-tosis through the downregulation of the HMGB1TLR4NF-120581B axis and autophagy through competitive interaction withBeclin 1 [46] In fact following 45min warm partial liver I-R injury in mice stressed hepatocytes released HMGB1 [46]When animals were treated with Ethyl-Pyruvate liver injuryapoptosis and necrosis were decreased as a result of HMGB1downregulation and autophagy was inhibited through thecompetitive interaction of HMGB1 with Beclin 1 [46]

Nevertheless a recent study suggested that HMGB1 isnot required for ATP production cellular respiration mito-chondrial architecture or autophagy in liver and heart [123]In mice with conditional HMGB1 deletion mitochondrialfunction and liver and heart autophagy were not affected[123] Other studies again outline the key role of HMGB1 inmitochondrial quality control and autophagy [124ndash126] Inparticular an aggravation of liver I-R injury after HMGB1depletion has been reported In fact genetic deletion ofHMGB1 from hepatocytes resulted in enhanced inflamma-tory signaling led to nuclear instability with increased DNAdamage and histone release led to mitochondria damage byexhausting nicotinamide adenine (NAD) and ATP storesexhibited increased ROS production and finally increasedcell death [126] In mice fasting for one day protected from60min warm liver I-R injury via Sirt1-dependent down-regulation of circulating HMGB1 [127] The reduced levels


of circulating HGMB1 damped the activation and self-prop-agation of Kupffer cells and hence protected from liver I-R[127]

33 Protective or Detrimental Role of Autophagy againstWarm andor Cold Liver I-R Injury Various Drugs andDifferent Methods for Monitoring Autophagy in DifferentAnimal and Human Experimental Liver Warm andor Cold I-R Models How pharmacological and surgical modulation ofliver autophagy could protect from or promote liver injuryfollowing I-R remains to be clarified as the studies on thissubject report either downregulated or excessive levels of liverautophagy At this point it is important to mention that themethods used for measuring autophagy were mainly steadystate methods the drugs used to modulate liver autophagywere not entirely specific for inhibition or stimulation ofautophagy and the animal and human experimental liver I-R models differed considerably In order to study the effectsof autophagy modulation on warm andor cold liver I-Rinjury both animal experimental liver I-Rmodels and humanexperimental I-R models are used The common length ofpartial warm ischemia in rodentmodelswas usually 30 45 60or 90min [128] In agreement with this as shown in Table 2in most of studies the role of autophagy on liver I-R injuryin rodents has been evaluated by using 60min partial lobar(70) liver warm ischemia model [28 32 37 42 44 47 48]The model of partial lobar (70) liver ischemia includesinterruption of blood flow to the left lateral and medianliver lobes leaving the right lobe for decompression [129]As animal studies overall mimic real clinic conditions inwhich liver surgery and liver transplantation are performedin humans they permit to draw conclusions with certainrelevance for the human physiopathology

331 Methods for Monitoring Autophagy Accumulationof autophagosomes may be due to both the induction ofautophagy and the blockage of a late step of the autophagyprocess including impaired autophagosome-lysosome fusionand compromised lysosomal activity [130] Induction ofautophagy assessed by steady state methods does not allowa determination of whether the autophagic process goes tocompletion [56] Incomplete autophagy which would leadto the accumulation of autophagosomes may contribute tocellular and organ dysfunction whereas complete autophagywill generally exert a cytoprotective effect [56] As steadystate methods evaluate autophagy only at a certain time point[56] they may not reflect properly the autophagic activity[56] Actually Table 2 shows that most studies on warmandor cold liver I-R [26 28ndash33 35ndash42 44 46] used steadystate methods electron microscopy Atg8LC3 western blot-ting and ubiquitin-like protein conjugation systems fluores-cence microscopy for monitoring phagophore and autoph-agosome formation

Electron Microscopy The autophagosome is a transientorganelle existing for less than 10min before fusing with thelysosome resulting in the appearance of autophagolysosomesat various stages of degradation [131] Electron microscopycan visualize early-stage autophagosomes but is less sensitive

for the visualization of late-stage autophagosomes [132]So the isolated approach with electron microscopy is notsufficient to evaluate autophagy levels [56]

LC3 Western Blotting LC3-II is present in most of theautophagic steps and reliably associated with phagophoressealed autophagosomes and mature autophagosomesautoly-sosomes [133] It is widely used to monitor autophagyImmunoblot analysis detects the conversion of LC3-I toLC3-II the amount of LC3-II is clearly correlated with thenumber of autophagosomes [134] However LC3-II itselfis degraded by autophagy and the amount of LC3-II at acertain time point does not necessarily indicate autoph-agic flux Simple comparison of LC3-I and LC3-II or sum-mation of LC3-II and LC3-II for ratio determination maynot be appropriate to correctly evaluate autophagy [134]An increased number of autophagosomes can occur despitelater steps of autophagy being blocked the quantifica-tion of LC3-II [134] before and after the inhibition of autoph-agosome-lysosome fusion by using lysosomal inhibitors mayindicate more accurately the autophagic flux [134] Chloro-quine and hydroxychloroquine increase the pH of the lyso-some Bafilomycin A1 inhibits the lysosomal Na+H+ ATPasein this way they prevent the activity of lysosomal acid pro-teases and cause autophagosomes to accumulate Similareffects are induced by treatment with specific inhibitors oflysosomal proteases such as Pepstatin A andor E64d [134]In this case the real autophagic flux is represented by thedifferent amounts of LC3-II in the samples in the absenceor presence of lysosomal proteases inhibitors LC3-II levelsproportionally increase in treated versus untreated samples[134]

Ubiquitin-Like Protein Conjugation Systems The p62(SQSTM1sequestosome 1) is a ubiquitin-binding scaffoldprotein that can bind LC3 [135] This protein accumulateswhen autophagy is inhibited and decreases when autophagyis induced [136] In some studies on warm or cold liver I-Rinjury in rats autophagy was monitored by degradation ofp62 using Western blot method [41 42 44] However p62is regulated at the transcriptional level by oxidative stressand by Ras oncogene and also feeds back to regulate NF-120581Bactivity [136] As p62 levels may be changed independentlyfrom autophagy additional methods to validate changesin protein aggregate turnover by autophagy are necessary[136ndash138]

Fluorescence MicroscopyThe fluorescent-based method withthe green fluorescent protein- (GFP-) LC3 counting the GFP-LC3 puncta uses the fact that after autophagy inductionLC3B becomes part of the newly formed autophagosomesand that GFP-LC3 changes its cellular localization from adiffuse cytosolic pattern to a punctate pattern Once againas steady state measurement this method is not sufficient tomeasure autophagy when used as an isolated approach [56]

Autophagy is a dynamic process of bulk degradationof cellular proteins and organelles in lysosomes [139]Autophagic substrates need to be monitored to verify thatthey have reached these organelles and eventually degraded


Table 2 Autophagy and warm andor cold liver I-R injury Methods for monitoring autophagy and additional beneficial effects of drugs andsurgical techniques on liver I-R injury other than modulation of autophagy

Authors andreferences

Monitoring autophagy byflux measurements [56]

Monitoring phagophore andautophagosome formation by steadystate methods [56]

Additional beneficial effects of drugs andsurgical techniques on liver I-R injury otherthan modulation of autophagy

Yun et al2014 [47] (+) HO-1 induction and Calpain 2 inhibition by

HeminKang et al2014 [48] (+) Decrease of apoptosis

Shen et al2013 [46] (+) Inhibits HMGB1TLR4NF-120581B axis inducing

apoptosis

Kim et al2013 [45]

(+) Suppression of calcium overloadingSuppression of uncontrolled Calpain activation

Fang et al2013 [44] (+)

Decrease of HMGB1 and proinflammatorycytokines levelsModulation of MAPK activation

Sun et al2013 [43] (+) Decreased mitochondrial ROS-inducing

necrosis by NAC

Liu et al2013 [42]

(+)Modulation of MAPK activationInhibition of Caspase-3 and -7 activationDecrease of HMGB1 and proinflammatorycytokines levels

Zaouali et al2013 [41] (+) Decrease of apoptosis

Gracia-Sanchoet al 2013 [40] (+) Decrease of the oxidative stress and Caspase-3

activation by Simvastatine

Bhogal et al2012 [39] (+)

Wang et al2012 [38] (+) Akt1 activation and decrease of apoptosis by

H2SEvankovichet al 2012 [37] (+)

Degli Espostiet al 2011 [35] (+)

Wang et al2011 [34] (+)

Minor et al2011 [36] (+) Decrease of HMGB1 and IFN beta levels

Minor et al2009 [30] (+) ROS decrease

ATP increase

Cardinal et al2009 [32] (+)


Gotoh et al2009 [33] (+)

Domart et al2009 [29] (+) Bcl-2 increase

Shin et al2008 [28] (+) ATP increase

Inhibition of Caspase-3 activationKim et al2008 [27] (+)

Cursio et al2010 [31] (+)

Lu et al2005 [26] (+)


[56] Evaluation of the autophagic flux a complete processof autophagy including the delivery of cargo to lysosomesvia its fusion with autophagosomes or amphisomes and itssubsequent breakdown and recycling [56] is important todetermine whether drugs andor surgical techniques trulyaffect autophagy As shown in Table 2 only a few studies havemonitored the autophagic flux in order to evaluate the extentof autophagy [27 34 43 45 47 48]

332 Side Effects of Chemical Autophagy Stimulators or Inhib-itors The chemical stimulators and inhibitors of autophagyused actually are not specific and may have a series of addi-tional effects on liver I-R apart from their action on auto-phagy

(1) Chemical Stimulators of Autophagy Inducing increasingor restoring basal autophagic activity in certain cell types asthe hepatocytes following warm andor cold liver I-R mightbe of therapeutic benefit As shown inTable 2 themodulationof autophagy by some stimulators protected against liver I-Rinjury [27 28 32 34 37 38 41 42 45] However the unspec-ificity of the used drugs renders the interpretation of theresults difficult

Rapamycin an autophagy inducer by inhibiting themTOR pathway plays a central role in several importantcellular processes other than autophagy [140] Interferingwith the translation of HIF-1120572 it has an antiangiogeniceffect and inhibiting the phosphorylation of BAD by S6K1Rapamycin may promote apoptosis [140] So its protectiveeffects in 90min warm liver I-R injury in mice may be partlydue to Akt1 activation and decreased apoptosis by H

2S asso-

ciated copretreatment [38] Similarly Rosiglitazone anotherautophagy stimulator has shown additional protective effects(increase of ATP levels and the inhibition of Caspase-3 acti-vation) in 30 60 and 90 min liver I-R injury in old mice thatcannot be accounted to the increase of autophagy [28]

The IMPase inhibitors Lithium chloride and Carba-mazepine can induce autophagy [92] They have also shownadditional protective effects against liver I-R injury otherthan autophagy induction [42 45] Lithium chloride showedmultiple additional effects the decrease of HMGB1 andproinflammatory cytokines levels the modulation of MAPKactivation and the inhibition of Caspase-3 and Caspase-7activation [42]The second Carbamazepine suppressed bothcalcium overloading and uncontrolled Calpain activation[45] In a murine model of 60 min liver I-R Cisplatin treat-ment increased liver autophagy and protected against I-Rinjury [32] Although the beneficial effects of Cisplatin are notonly the result of autophagy stimulation but also of decreasedHMGB1 and proinflammatory cytokines levels and of themodulation of MAPK activation [32]

In steatotic rat livers that were preserved in cold solutionfor 24 hours autophagy was decreased [41] A close relation-ship between 51015840 AMP-activated protein kinase (AMPK) acti-vation and endoplasmic reticulum (ER) stress and autophagyhas been observed in these livers [41]The addition of aMela-tonin and Trimetazidine cocktail to the cold preservationsolution improved steatotic liver graft preservation throughAMPK activation which in turn reduced ER stress and

increased autophagy [41] However these beneficial effectsof the Melatonin and Trimetazidine cocktail may be due inpart to a reduction of apoptotic liver cells mainly observed inperiportal and midzonal areas of the liver [41]

In human liver surgery and transplantation the improve-ment of clinical outcomes by the use of IP of the liverwas associated to higher levels of autophagy However thedecrease of ER stress [35] and the increase of ATP andBeclin 1 levels observed after IP of the liver may have beenresponsible too for the beneficial effects of this surgicalstrategy [29]

(2) Chemical Inhibitors of Autophagy Similar to autophagystimulators autophagy inhibitors are not specific [33 3943 44] and additional effects of these drugs may play animportant role [43 44 46ndash48]

Autophagy inhibition by 3-MA in human hypoxemicreoxygenated hepatocytes resulted in MPT lowering andonset of apoptosis [39] Autophagy inhibition by LY294002andWortmannin during liver transplantation in rats reducedliver graft dysfunction and mortality rate of transplantedanimals [33]

The PI3K inhibitors 3-MA LY294002 andWortmannintypically block Class III PI3Ks which act as downstreamof the negative regulatory Class I kinase However theirinhibitory action on autophagy differs [141] In fact 3-MAmay promote or suppress autophagic flux [142] whereasWortmannin seems to have inhibitory effects opposite tothose of 3-MA It has persistent effects on class III PI3K anautophagy activator but also it has transient effects on class IPI3K which is an autophagy inhibitor [142] The autophagicinhibitor LY294002 plays also a dual role in the regulation ofautophagy as itmay activate autophagy by inhibiting the classI PI3K [143] and as itmodulates calciumoverloading Calpainactivation and MPT all implicated in the development ofwarm liver I-R injury [77 81]

Also Melatonin Ethyl-Pyruvate and Chloroquine allthree autophagy inhibitors present additional protectiveeffects against liver I-R injuryMelatonin in fact may decreaseliver apoptosis [48] Ethyl-Pyruvate may inhibit HMGB1TLR4NF-120581B axis inducing apoptosis [46] and Chloroquinewhich has protective effects in the early reperfusion phaseafter 60 and 90min warm liver ischemia may modulateMAPK activation [44]

Autophagy inhibition by Chloroquine pretreatmentincreased mitochondrial oxidative stress and hepatocellularnecrosis following 90min warm liver I-R in rats [43]Antioxidant NAC pretreatment again diminished the ische-mia-induced liver injury of these rats which received alsoChloroquine treatment However the beneficial effectsagainst liver injury by NAC seem to be due also to decreasedmitochondrial ROS-inducing necrosis [43]

4 Conclusion

The pathogenesis of warm andor cold liver I-R injuryrepresents a complex interplay between necrosis apopto-sis and autophagy It seems that stimulation of autophagyplays a more important role during liver reperfusion than


ischemia Depending on the context induction or impair-ment of autophagy during warm andor cold liver I-R canbe protective or detrimental for liver cells Stimulation ofimpaired autophagy following warm andor cold I-R maypromote hepatocyte survival by degradation of intracellu-lar contents to maintain ATP production and removal ofdamaged organelles and protein aggregates Excessive andlong-term upregulation of autophagy as it occurs duringsevere ischemic insult of the liver may lead to destruction ofessential proteins and organelles resulting in hepatocellularapoptosis and necrosis

However the results of the studies on autophagy duringwarm andor cold liver I-R remain discordant This may bedue to several factors namely the lack of drugs which exertspecific and exclusive autophagic stimulation or inhibitionthe different experimental liver I-R models used and thedifferent methods of autophagy evaluation So how pharma-cological andor surgicalmodulation of liver autophagy couldprotect from or promote liver injury following warm andorcold I-R remains to be clarified Large animal studies onliver I-R also at a genetic level with knockout models whichprovide a very specific targeted disruption of a particularautophagic protein and will therefore be more informativethan the use of not entirely specific chemical stimulators orinhibitors of the same autophagic protein are needed Lastbut not least the methods for monitoring autophagy shouldpreferentially measure the autophagic flux

In any way the autophagic cell response to warm andorcold liver I-R may provide an additional time to the celldeath processes delaying apoptosis and necrosis and thusultimately increasing the possibility for novel therapeuticintervention to diminish the extent of warmcold liver I-Rinjury

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] H Jaeschke ldquoMechanisms of reperfusion injury after warmischemia of the liverrdquo Journal of Hepato-Biliary-PancreaticSurgery vol 5 no 4 pp 402ndash408 1998

[2] R Cursio ldquoCaspase inhibition in liver transplantation frombasic research to clinical studiesrdquo HPB vol 12 no 1 pp 1ndash32010

[3] H Jaeschke ldquoMolecularmechanisms of hepatic ischemia-reper-fusion injury and preconditioningrdquo American Journal of Phys-iologymdashGastrointestinal and Liver Physiology vol 284 no 1 ppG15ndashG26 2003

[4] R Cursio Liver ischemia-reperfusion injury in the rat role ofnonparenchymal liver cells [Postdoctoral thesis on liver transplan-tation] University of Paris XI Paris France 2002

[5] G C Velmahos K Toutouzas R Radin et al ldquoHigh successwith nonoperative management of blunt hepatic trauma theliver is a sturdy organrdquo Archives of Surgery vol 138 no 5 pp475ndash481 2003

[6] H T Hassoun B C Kone D W Mercer F G Moody N WWeisbrodt and F AMoore ldquoPost-injurymultiple organ failurethe role of the gutrdquo Shock vol 15 no 1 pp 1ndash10 2001

[7] J Belghiti K Hiramatsu S Benoist P P Massault A SauvanetandO Farges ldquoSeven hundred forty-seven hepatectomies in the1990s an update to evaluate the actual risk of liver resectionrdquoJournal of the American College of Surgeons vol 191 no 1 pp38ndash46 2000

[8] P-A Clavien M Selzner H A Rudiger et al ldquoA prospectiverandomized study in 100 consecutive patients undergoingmajor liver resection with versus without ischemic precondi-tioningrdquo Annals of Surgery vol 238 no 6 pp 843ndash852 2003

[9] P-A Clavien P R CHarvey and SM Strasberg ldquoPreservationand reperfusion injuries in liver allografts an overview andsynthesis of current studiesrdquo Transplantation vol 53 no 5 pp957ndash978 1992

[10] D G Farmer F Amersi J Kupiec-Weglinski and RW BusuttilldquoCurrent status of ischemia and reperfusion injury in the liverrdquoTransplantation Reviews vol 14 no 2 pp 106ndash126 2000

[11] C Fondevila R W Busuttil and J W Kupiec-Weglinski ldquoHep-atic ischemia-reperfusion injurymdasha fresh lookrdquo Experimentaland Molecular Pathology vol 74 no 2 pp 86ndash93 2003

[12] M Glanemann J M Langrehr B J Stange et al ldquoClinicalimplications of hepatic preservation injury after adult livertransplantationrdquo The American Journal of Transplantation vol3 no 8 pp 1003ndash1009 2003

[13] O de Rougemont K Lehmann and P-A Clavien ldquoPrecondi-tioning organ preservation and postconditioning to preventischemia-reperfusion injury to the liverrdquo Liver Transplantationvol 15 no 10 pp 1172ndash1182 2009

[14] L Sanchez-Urdazpal G J Gores E M Ward et al ldquoIschemic-type biliary complications after orthotopic liver transplanta-tionrdquo Hepatology vol 16 no 1 pp 49ndash53 1992

[15] C I Buis H Hoekstra R C Verdonk and R J Porte ldquoCausesand consequences of ischemic-type biliary lesions after livertransplantationrdquo Journal of Hepato-Biliary-Pancreatic Surgeryvol 13 no 6 pp 517ndash524 2006

[16] R Cursio and J Gugenheim ldquoIschemia-reperfusion injury andischemic-type biliary lesions following liver transplantationrdquoJournal of Transplantation vol 2012 Article ID 164329 17 pages2012

[17] P FHalloran ldquoCall for revolution a new approach to describingallograft deteriorationrdquo American Journal of Transplantationvol 2 no 3 pp 195ndash200 2002

[18] S Gourishankar and P F Halloran ldquoLate deterioration of organtransplants a problem in injury and homeostasisrdquo CurrentOpinion in Immunology vol 14 no 5 pp 576ndash583 2002

[19] P-A Clavien ldquoHow far can we go with marginal donorsrdquo Jour-nal of Hepatology vol 45 no 4 pp 483ndash484 2006

[20] N Selzner H Rudiger R Graf and P-A Clavien ldquoProtectivestrategies against ischemic injury of the liverrdquoGastroenterologyvol 125 no 3 pp 917ndash936 2003

[21] H Jaeschke and J J Lemasters ldquoApoptosis versus oncotic necro-sis in hepatic ischemiareperfusion injuryrdquo Gastroenterologyvol 125 no 4 pp 1246ndash1257 2003

[22] J J Lemasters A-L Nieminen T Qian et al ldquoThe mito-chondrial permeability transition in cell death a commonmechanism in necrosis apoptosis and autophagyrdquo Biochimicaet Biophysica Acta vol 1366 no 1-2 pp 177ndash196 1998

[23] J J Lemasters ldquoV Necrapoptosis and themitochondrial perme-ability transition shared pathways to necrosis and apoptosisrdquo


The American Journal of Physiology vol 276 no 1 part 1 ppG1ndashG6 1999

[24] R Cursio J Gugenheim J E Ricci et al ldquoA caspase inhibitorfully protects rats against lethal normothermic liver ischemiaby inhibition of liver apoptosisrdquoThe FASEB Journal vol 13 no2 pp 253ndash261 1999

[25] W Gao R C Bentley J E Madden and P-A ClavienldquoApoptosis of sinusoidal endothelial cells is a criticalmechanismof preservation injury in rat liver transplantationrdquo Hepatologyvol 27 no 6 pp 1652ndash1660 1998

[26] Z Lu K Dono K Gotoh et al ldquoParticipation of autophagyin the degeneration process of rat hepatocytes after trans-plantation following prolonged cold preservationrdquo Archives ofHistology and Cytology vol 68 no 1 pp 71ndash80 2005

[27] J-S Kim T Nitta D Mohuczy et al ldquoImpaired autophagy amechanism of mitochondrial dysfunction in anoxic rat hepato-cytesrdquo Hepatology vol 47 no 5 pp 1725ndash1736 2008

[28] T Shin S Kuboki N Huber et al ldquoActivation of peroxisomeproliferator-activated receptor-120574during hepatic ischemia is age-dependentrdquo Journal of Surgical Research vol 147 no 2 pp 200ndash205 2008

[29] M-C Domart D D Esposti M Sebagh et al ldquoConcurrentinduction of necrosis apoptosis and autophagy in ischemicpreconditioned human livers formerly treated by chemother-apyrdquo Journal of Hepatology vol 51 no 5 pp 881ndash889 2009

[30] T Minor J Stegemann A Hirner and M Koetting ldquoImpairedautophagic clearance after cold preservation of fatty liverscorrelates with tissue necrosis upon reperfusion and is reversedby hypothermic reconditioningrdquo Liver Transplantation vol 15no 7 pp 798ndash805 2009

[31] R Cursio P Colosetti M-C Saint-Paul et al ldquoInduction ofdifferent types of cell death after normothermic liver ischemia-reperfusionrdquo Transplantation Proceedings vol 42 no 10 pp3977ndash3980 2010

[32] J Cardinal P Pan M Ross et al ldquoCisplatin prevents highmobility group box 1 release and is protective in amurinemodelof hepatic ischemiareperfusion injuryrdquoHepatology vol 50 no2 pp 565ndash574 2009

[33] K Gotoh Z Lu MMorita et al ldquoParticipation of autophagy inthe initiation of graft dysfunction after rat liver transplantationrdquoAutophagy vol 5 no 3 pp 351ndash360 2009

[34] J Wang I Ahn T D Fischer et al ldquoAutophagy suppresses age-dependent ischemia and reperfusion injury in livers of micerdquoGastroenterology vol 141 no 6 pp 2188e6ndash2199e6 2011

[35] D Degli Esposti M Sebagh P Pham et al ldquoIschemic pre-conditioning induces autophagy and limits necrosis in humanrecipients of fatty liver grafts decreasing the incidence ofrejection episodesrdquo Cell Death and Disease vol 2 no 1 articlee111 2011

[36] T Minor M Koetting G Kaiser P Efferz B Luer and A PaulldquoHypothermic reconditioning by gaseous oxygen improvessurvival after liver transplantation in the pigrdquoAmerican Journalof Transplantation vol 11 no 12 pp 2627ndash2634 2011

[37] J Evankovich R Zhang J S Cardinal et al ldquoCalciumcal-modulin-dependent protein kinase IV limits organ damagein hepatic ischemia-reperfusion injury through induction ofautophagyrdquo American Journal of PhysiologymdashGastrointestinaland Liver Physiology vol 303 no 2 pp G189ndashG198 2012

[38] D Wang Y Ma Z Li et al ldquoThe role of AKT1 and autophagyin the protective effect of hydrogen sulphide against hepaticischemiareperfusion injury in micerdquo Autophagy vol 8 no 6pp 954ndash962 2012

[39] R H Bhogal C J Weston S M Curbishley D H Adams andS C Afford ldquoAutophagy a cyto-protective mechanism whichprevents primary human hepatocyte apoptosis during oxidativestressrdquo Autophagy vol 8 no 4 pp 545ndash558 2012

[40] J Gracia-Sancho H Garcıa-Caldero D Hide et al ldquoSimvas-tatin maintains function and viability of steatotic rat liversprocured for transplantationrdquo Journal of Hepatology vol 58 no6 pp 1140ndash1146 2013

[41] M A Zaouali E Boncompagni R J Reiter et al ldquoAMPKinvolvement in endoplasmic reticulum stress and autophagymodulation after fatty liver graft preservation a role for mela-tonin and trimetazidine cocktailrdquo Journal of Pineal Researchvol 55 no 1 pp 65ndash78 2013

[42] A Liu H Fang U Dahmen and O Dirsch ldquoChronic lithiumtreatment protects against liver ischemiareperfusion injury inratsrdquo Liver Transplantation vol 19 no 7 pp 762ndash772 2013

[43] K Sun X Xie Y Liu et al ldquoAutophagy lessens ischemic liverinjury by reducing oxidative damagerdquo Cell amp Bioscience vol 3article 26 2013

[44] H Fang A Liu U Dahmen and O Dirsch ldquoDual role ofchloroquine in liver ischemia reperfusion injury reduction ofliver damage in early phase but aggravation in late phaserdquo CellDeath amp Disease vol 4 article e694 2013

[45] J-S Kim J-H Wang T G Biel et al ldquoCarbamazepine sup-presses calpain-mediated autophagy impairment after ische-miareperfusion in mouse liversrdquo Toxicology and Applied Phar-macology vol 273 no 3 pp 600ndash610 2013

[46] M Shen J LuW Dai et al ldquoEthyl pyruvate ameliorates hepaticischemia-reperfusion injury by inhibiting intrinsic pathway ofapoptosis and autophagyrdquoMediators of Inflammation vol 2013Article ID 461536 12 pages 2013

[47] N Yun H-I Cho and S-M Lee ldquoImpaired autophagy con-tributes to hepatocellular damage during ischemiareperfusionheme oxygenase-1 as a possible regulatorrdquo Free Radical Biologyand Medicine vol 68 pp 168ndash177 2014

[48] J-W Kang H-I Cho and S-M Lee ldquoMelatonin inhibitsmTOR-dependent autophagy during liver ischemiareperfu-sionrdquoCellular Physiology and Biochemistry vol 33 no 1 pp 23ndash36 2014

[49] E H Baehrecke ldquoAutophagy dual roles in life and deathrdquoNature Reviews Molecular Cell Biology vol 6 no 6 pp 505ndash510 2005

[50] D J Klionsky ldquoThe molecular machinery of autophagy unan-swered questionsrdquo Journal of Cell Science vol 118 no 1 pp 7ndash182005

[51] Y Chen and D J Klionsky ldquoThe regulation of autophagymdashunanswered questionsrdquo Journal of Cell Science vol 124 no 2pp 161ndash170 2011

[52] P-E Rautou A Mansouri D Lebrec F Durand D Valla andR Moreau ldquoAutophagy in liver diseasesrdquo Journal of Hepatologyvol 53 no 6 pp 1123ndash1134 2010

[53] P Codogno and A J Meijer ldquoAutophagy in the liverrdquo Journal ofHepatology vol 59 no 2 pp 389ndash391 2013

[54] M J Czaja W-X Ding T M Donohue Jr et al ldquoFunctions ofautophagy in normal and diseased liverrdquo Autophagy vol 9 no8 pp 1131ndash1158 2013

[55] X-M Yin W-X Ding and W Gao ldquoAutophagy in the liverrdquoHepatology vol 47 no 5 pp 1773ndash1785 2008

[56] D J Klionski H Abeliovich P Agostinis D K Agrawal andG Aliev ldquoGuidelines for the use and interpretation of assays formonitoring autophagy in higher eukaryotesrdquo Autophagy vol 4no 2 pp 151ndash175 2008


[57] R GThurman I Marzi G Seitz J Thies J J Lemasters and FZimmerman ldquoHepatic reperfusion injury following orthotopicliver transplantation in the ratrdquo Transplantation vol 46 no 4pp 502ndash506 1988

[58] C M B McKeown V Edwards M J Phillips P R C HarveyC N Petrunka and S M Strasberg ldquoSinusoidal lining celldamage the critical injury in cold preservation of liver allograftsin the ratrdquo Transplantation vol 46 no 2 pp 178ndash191 1988

[59] J C Caldwell-Kenkel R T Currin Y Tanaka R G Thurmanand J J Lemasters ldquoReperfusion injury to endothelial cellsfollowing ischemic storage of rat liversrdquoHepatology vol 10 no3 pp 292ndash299 1989

[60] M Mendes-Braz M Elias-Miro M B Jimenez-Castro ACasillas-Ramırez F S Ramalho and C Peralta ldquoThe currentstate of knowledge of hepatic ischemia-reperfusion injury basedon its study in experimentalmodelsrdquo Journal of Biomedicine andBiotechnology vol 2012 Article ID 298657 20 pages 2012

[61] J J Lemasters and R G Thurman ldquoReperfusion injury afterliver preservation for transplantationrdquo Annual Review of Phar-macology and Toxicology vol 37 pp 327ndash338 1997

[62] E E Montalvo-Jave T Escalante-Tattersfield J A Ortega-Salgado E Pina and D A Geller ldquoFactors in the pathophysiol-ogy of the liver ischemia-reperfusion injuryrdquo Journal of SurgicalResearch vol 147 no 1 pp 153ndash159 2008

[63] H Jaeschke A Farhood A P Bautista Z Spolarics and J JSpitzer ldquoComplement activates Kupffer cells and neutrophilsduring reperfusion after hepatic ischemiardquo American Journal ofPhysiologymdashGastrointestinal and Liver Physiology vol 264 no4 part 1 pp G801ndashG809 1993

[64] H Jaeschke ldquoMechanisms of liver injury II Mechanisms ofneutrophil-induced liver cell injury during hepatic ischemia-reperfusion and other acute inflammatory conditionsrdquo TheAmerican Journal of PhysiologymdashGastrointestinal and LiverPhysiology vol 290 no 6 pp G1083ndashG1088 2006

[65] N A Essani G M McGuire A M Manning and H JaeschkeldquoEndotoxin-induced activation of the nuclear transcriptionfactor 120581B and expression of E-selectin messenger RNA inhepatocytes kupffer cells and endothelial cells in vivordquo TheJournal of Immunology vol 156 no 8 pp 2956ndash2963 1996

[66] T Matsumura A Ito T Takii H Hayashi and K OnozakildquoEndotoxin and cytokine regulation of toll-like receptor (TLR)2 and TLR4 gene expression in murine liver and hepatocytesrdquoJournal of Interferon and Cytokine Research vol 20 no 10 pp915ndash921 2000

[67] N C Teoh ldquoHepatic ischemia reperfusion injury contem-porary perspectives on pathogenic mechanisms and basis forhepatoprotection-the good bad and deadlyrdquo Journal of Gas-troenterology andHepatology vol 26 supplement 1 pp 180ndash1872011

[68] B Vollmar J Glasz R Leiderer S Post and M D MengerldquoHepatic microcirculatory perfusion failure is a determinant ofliver dysfunction in warm ischemia-reperfusionrdquoTheAmericanJournal of Pathology vol 145 no 6 pp 1421ndash1431 1994

[69] W Zhou M O McCollum B A Levine and M S OlsonldquoInflammation and platelet-activating factor production duringhepatic ischemiareperfusionrdquo Hepatology vol 16 no 5 pp1236ndash1240 1992

[70] X Qin and B Gao ldquoThe complement system in liver diseasesrdquoCellular amp Molecular Immunology vol 3 no 5 pp 333ndash3402006

[71] P Boros and J S Bromberg ldquoNew cellular and molecularimmune pathways in ischemiareperfusion injuryrdquo The Amer-ican Journal of Transplantation vol 6 no 4 pp 652ndash658 2006

[72] Y Zhai R W Busuttil and J W Kupiec-Weglinski ldquoLiverischemia and reperfusion injury new insights intomechanismsof innate-adaptive immune-mediated tissue inflammationrdquoAmerican Journal of Transplantation vol 11 no 8 pp 1563ndash1569 2011

[73] Y Zhai H Petrowsky J C Hong R W Busuttil and JW Kupiec-Weglinski ldquoIschaemia-reperfusion injury in livertransplantation-from bench to bedsiderdquo Nature Reviews Gas-troenterology and Hepatology vol 10 no 2 pp 79ndash89 2013

[74] Y Huang H Rabb and K L Womer ldquoIschemia-reperfusionand immediate T cell responsesrdquoCellular Immunology vol 248no 1 pp 4ndash11 2007

[75] D Linfert T Chowdhry and H Rabb ldquoLymphocytes andischemia-reperfusion injuryrdquo Transplantation Reviews vol 23no 1 pp 1ndash10 2009

[76] C C Caldwell T Okaya A Martignoni T Husted R Schusterand A B Lentsch ldquoDivergent functions of CD4+ T lympho-cytes in acute liver inflammation and injury after ischemia-reperfusionrdquo American Journal of PhysiologymdashGastrointestinaland Liver Physiology vol 289 no 5 pp G969ndashG976 2005

[77] J-S Kim T Qian and J J Lemasters ldquoMitochondrial perme-ability transition in the switch from necrotic to apoptotic celldeath in ischemic rat hepatocytesrdquo Gastroenterology vol 124no 2 pp 494ndash503 2003

[78] R Cursio P Colosetti P Auberger and J Gugenheim ldquoAuto-phagy and liver ischemia-reperfusion injuryrdquo in AutophagyPrinciples Regulation and Role in Diseases pp 297ndash328 NovaNew York NY USA 2012

[79] Z Xie and D J Klionsky ldquoAutophagosome formation coremachinery and adaptationsrdquo Nature Cell Biology vol 9 no 10pp 1102ndash1109 2007

[80] R Kang H J Zeh M T Lotze and D Tang ldquoThe Beclin 1network regulates autophagy and apoptosisrdquo Cell Death andDifferentiation vol 18 no 4 pp 571ndash580 2011

[81] S A Susin H K Lorenzo N Zamzami et al ldquoMolecular char-acterization ofmitochodrial apoptosis-inducing factorrdquoNaturevol 397 no 6718 pp 441ndash446 1999

[82] S P Elmore T Qian S F Grissom and J J Lemasters ldquoThemitochondrial permeability transition initiates autophagy in rathepatocytesrdquoThe FASEB Journal vol 15 no 12 pp 2286ndash22872001

[83] J R Klune and A Tsung ldquoMolecular biology of liver ische-miareperfusion injury established mechanisms and recentadvancementsrdquo Surgical Clinics of North America vol 90 no4 pp 665ndash677 2010

[84] I Ito J Fukazawa and M Yoshida ldquoPost-translational methy-lation of high mobility group box 1 (HMGB1) causes its cyto-plasmic localization in neutrophilsrdquo The Journal of BiologicalChemistry vol 282 no 22 pp 16336ndash16344 2007

[85] P Scaffidi T Misteli and M E Bianchi ldquoRelease of chro-matin protein HMGB1 by necrotic cells triggers inflammationrdquoNature vol 418 no 6894 pp 191ndash195 2002

[86] S Muller P Scaffidi B Degryse et al ldquoThe double life ofHMGB1 chromatin protein architectural factor and extracellu-lar signalrdquo The EMBO Journal vol 20 no 16 pp 4337ndash43402001


[87] A Tsung R Sahai H Tanaka et al ldquoThe nuclear factorHMGB1 mediates hepatic injury after murine liver ischemia-reperfusionrdquo Journal of Experimental Medicine vol 201 no 7pp 1135ndash1143 2005

[88] A Liu H Jin O Dirsch et al ldquoRelease of danger signals dur-ing ischemic storage of the liver a potential marker of organdamagerdquo Mediators of Inflammation vol 2010 Article ID436145 11 pages 2010

[89] A Liu H Fang O Dirsch H Jin and U Dahmen ldquoOxidationof hmgb1 causes attenuation of its pro-inflammatory activityand occurs during liver ischemia and reperfusionrdquo PLoS ONEvol 7 no 4 Article ID e35379 2012

[90] A Liu O Dirsch H Fang et al ldquoHMGB1 in ischemic and non-ischemic liver after selective warm ischemiareperfusion in ratrdquoHistochemistry and Cell Biology vol 135 no 5 pp 443ndash4522011

[91] M Ilmakunnas E M Tukiainen A Rouhiainen et al ldquoHighmobility group box 1 protein as a marker of hepatocellularinjury in human liver transplantationrdquo Liver Transplantationvol 14 no 10 pp 1517ndash1525 2008

[92] S Sarkar R A Floto Z Berger et al ldquoLithium induces auto-phagy by inhibiting inositol monophosphataserdquo The Journal ofCell Biology vol 170 no 7 pp 1101ndash1111 2005

[93] C J Mullonkal and L H Toledo-Pereyra ldquoAkt in ischemia andreperfusionrdquo Journal of Investigative Surgery vol 20 no 3 pp195ndash203 2007

[94] C Muller F Dunschede E Koch A M Vollmar and AK Kiemer ldquo120572-lipoic acid preconditioning reduces ischemia-reperfusion injury of the rat liver via the PI3-kinaseAktpathwayrdquoThe American Journal of PhysiologymdashGastrointestinaland Liver Physiology vol 285 no 4 pp G769ndashG778 2003

[95] R Cursio N Filippa C Miele E Van Obberghen and JGugenheim ldquoInvolvement of protein kinase B and mitogen-activated protein kinases in experimental normothermic liverischaemia-reperfusion injuryrdquo The British Journal of Surgeryvol 93 no 6 pp 752ndash761 2006

[96] X L Li KManK TNg C K Sun CM Lo and S T Fan ldquoTheinfluence of phosphatidylinositol 3-kinaseAkt pathway on theischemic injury during rat liver graft preservationrdquo AmericanJournal of Transplantation vol 5 no 6 pp 1264ndash1275 2005

[97] Y Kurihara T Kanki Y Aoki et al ldquoMitophagy plays anessential role in reducing mitochondrial production of reactiveoxygen species and mutation of mitochondrial DNA by main-tainingmitochondrial quantity andquality in yeastrdquoTheJournalof Biological Chemistry vol 287 no 5 pp 3265ndash3272 2012

[98] AM Cuervo E Bergamini U T BrunkW DrogeM FfrenchandA Terman ldquoAutophagy and aging the importance ofmain-taining ldquocleanrdquo cellsrdquo Autophagy vol 1 no 3 pp 131ndash140 2005

[99] S Kuboki T Shin N Huber et al ldquoPeroxisome proliferator-activated receptor-120574 protects against hepatic ischemiareper-fusion injury in micerdquo Hepatology vol 47 no 1 pp 215ndash2242008

[100] V E de Meijer B T Kalish M Puder and J N M IJzermansldquoSystematic review andmeta-analysis of steatosis as a risk factorinmajor hepatic resectionrdquoBritish Journal of Surgery vol 97 no9 pp 1331ndash1339 2010

[101] D J Verran T Kusyk D Painter et al ldquoClinical experiencegained from the use of 120 steatotic donor livers for orthotopicliver transplantationrdquo Liver Transplantation vol 9 no 5 pp500ndash505 2003

[102] P Caraceni C Bianchi M Domenicali et al ldquoImpairmentof mitochondrial oxidative phosphorylation in rat fatty liver

exposed to preservation-reperfusion injuryrdquo Journal of Hepa-tology vol 41 no 1 pp 82ndash88 2004

[103] M J J Chu A J R Hickey A R J Phillips and A S J RBartlett ldquoThe impact of hepatic steatosis on hepatic ischemia-reperfusion injury in experimental studies a systematic reviewrdquoBioMed Research International vol 2013 Article ID 192029 12pages 2013

[104] Y Wang R Singh Y Xiang and M J Czaja ldquoMacroautophagyand chaperone-mediated autophagy are required for hepatocyteresistance to oxidant stressrdquo Hepatology vol 52 no 1 pp 266ndash277 2010

[105] R Singh S Kaushik Y Wang et al ldquoAutophagy regulates lipidmetabolismrdquo Nature vol 458 no 7242 pp 1131ndash1135 2009

[106] T Minor S Akbar R Tolba and F Dombrowski ldquoCold pres-ervation of fatty liver grafts prevention of functional andultrastructural impairments by venous oxygen persufflationrdquoJournal of Hepatology vol 32 no 1 pp 105ndash111 2000

[107] T Minor and M Kotting ldquoGaseous oxygen for hypothermicpreservation of predamaged liver grafts fuel to cellular home-ostasis or radical tissue alterationrdquo Cryobiology vol 40 no 2pp 182ndash186 2000

[108] R Scherz-Shouval Y Sagiv H Shorer and Z Elazar ldquoTheCOOH terminus of GATE-16 an intra-golgi transport modu-lator is cleaved by the human cysteine protease HsApg4ArdquoTheJournal of Biological Chemistry vol 278 no 16 pp 14053ndash140582003

[109] C Peralta G Hotter D Closa E Gelpı O Bulbena and JRosello-Catafau ldquoProtective effect of preconditioning on theinjury associated to hepatic ischemia-reperfusion in the rat roleof nitric oxide and adenosinerdquo Hepatology vol 25 no 4 pp934ndash937 1997

[110] P-A Clavien S Yadav D Sindram and R C Bentley ldquoPro-tective effects of ischemic preconditioning for liver resectionperformed under inflow occlusion in humansrdquo Annals ofSurgery vol 232 no 2 pp 155ndash162 2000

[111] H Petrowsky L McCormack M Trujillo M Selzner WJochum and P-A Clavien ldquoA prospective randomized con-trolled trial comparing intermittent portal triad clamping ver-sus ischemic preconditioning with continuous clamping formajor liver resectionrdquoAnnals of Surgery vol 244 no 6 pp 921ndash928 2006

[112] D Azoulay M Del Gaudio P Andreani et al ldquoEffects of 10minutes of ischemic preconditioning of the cadaveric liver onthe graftrsquos preservation and function the Ying and the YangrdquoAnnals of Surgery vol 242 no 1 pp 133ndash139 2005

[113] K S Gurusamy Y Kumar D Sharma and B R David-son ldquoIschaemic preconditioning for liver transplantationrdquo TheCochrane Database of Systematic Reviews vol 21 no 1 ArticleID CD006315 2008

[114] N Selzner M Boehnert and M Selzner ldquoPreconditioningpostconditioning and remote conditioning in solid organtransplantation basic mechanisms and translational applica-tionsrdquo Transplantation Reviews vol 26 no 2 pp 115ndash124 2012

[115] S OrsquoNeill S Leuschner S J McNally O J Garden S JWigmore and E M Harrison ldquoMeta-analysis of ischaemicpreconditioning for liver resectionsrdquo The British Journal ofSurgery vol 100 no 13 pp 1689ndash1700 2013

[116] F G Fernandez J Ritter J W Goodwin D C LinehanW G Hawkins and S M Strasberg ldquoEffect of steatohepati-tis associated with irinotecan or oxaliplatin pretreatment onresectability of hepatic colorectal metastasesrdquo Journal of theAmerican College of Surgeons vol 200 no 6 pp 845ndash853 2005


[117] K E Behrns G G Tsiotos N F DeSouza M K Krishna JLudwig and D M Nagorney ldquoHepatic steatosis as a potentialrisk factor for major hepatic resectionrdquo Journal of Gastrointesti-nal Surgery vol 2 no 3 pp 292ndash298 1998

[118] Y Matsui H Takagi X Qu et al ldquoDistinct roles of autophagyin the heart during ischemia and reperfusion roles of AMP-activated protein kinase and beclin 1 in mediating autophagyrdquoCirculation Research vol 100 no 6 pp 914ndash922 2007

[119] M Vairetti A Ferrigno R Bertone et al ldquoExogenous mela-tonin enhances bile flow and ATP levels after cold storage andreperfusion in rat liver implications for liver transplantationrdquoJournal of Pineal Research vol 38 no 4 pp 223ndash230 2005

[120] I Freitas V Bertone C Guarnaschelli et al ldquoIn situ demon-stration of improvement of liver mitochondria function bymelatonin after cold ischemiardquo In Vivo vol 20 no 2 pp 229ndash237 2006

[121] L Russo J Gracia-Sancho H Garcıa-Caldero et al ldquoAdditionof simvastatin to cold storage solution prevents endothelialdysfunction in explanted rat liversrdquo Hepatology vol 55 no 3pp 921ndash930 2012

[122] R Kang K M Livesey H J Zeh M T Lotze and D TangldquoHMGB1 a novel Beclin 1-binding protein active in autophagyrdquoAutophagy vol 6 no 8 pp 1209ndash1211 2010

[123] P PHuebenerG-E YGwak J-P Pradere et al ldquoHigh-mobilitygroup box 1 is dispensable for autophagy mitochondrial qualitycontrol and organ function in vivordquoCellMetabolism vol 19 pp539ndash547 2014

[124] D Tang R Kang KM Livesey et al ldquoHigh-mobility group box1 is essential formitochondrial quality controlrdquoCellMetabolismvol 13 no 6 pp 701ndash711 2011

[125] R Kang K M Livesey H J Zeh M T Lotze and D TangldquoMetabolic regulation by HMGB1-mediated autophagy andmitophagyrdquo Autophagy vol 7 no 10 pp 1256ndash1258 2011

[126] H Huang G W Nace K-A Mcdonald et al ldquoHepatocyte-specific high-mobility group box 1 deletion worsens the injuryin liver ischemiareperfusion a role for intracellular high-mobility group box 1 in cellular protectionrdquoHepatology vol 59no 5 pp 1984ndash1997 2014

[127] A Rickenbacher J H Jang P Limani et al ldquoFasting protectsliver from ischemic injury through Sirt1-mediated downregula-tion of circulating HMGB1 in micerdquo Journal of Hepatology vol61 no 2 pp 301ndash308 2014

[128] T Karatzas A-A Neri M-E Baibaki and I A DontasldquoRodent models of hepatic ischemia-reperfusion injury timeand percentage-related pathophysiological mechanismsrdquo Jour-nal of Surgical Research vol 191 no 2 pp 399ndash412 2014

[129] HU Spiegel andR Bahde ldquoExperimentalmodels of temporarynormothermic liver ischemiardquo Journal of Investigative Surgeryvol 19 no 2 pp 113ndash123 2006

[130] N Mizushima T Yoshimori and B Levine ldquoMethods in mam-malian autophagy researchrdquo Cell vol 140 no 3 pp 313ndash3262010

[131] Y Ohsumi ldquoHistorical landmarks of autophagy researchrdquo CellResearch vol 24 no 1 pp 9ndash23 2014

[132] J M Swanlund K C Kregel and T D Oberley ldquoInvestigatingautophagy quantitative morphometric analysis using electronmicroscopyrdquo Autophagy vol 6 no 2 pp 270ndash277 2010

[133] D C Rubinsztein A M Cuervo B Ravikumar et al ldquoIn searchof an ldquoautophagomometerrdquordquo Autophagy vol 5 no 5 pp 585ndash589 2009

[134] N Mizushima and T Yoshimori ldquoHow to interpret LC3immunoblottingrdquo Autophagy vol 3 no 6 pp 542ndash545 2007

[135] M L Seibenhener T Geetha and M W Wooten ldquoSequesto-some 1p62mdashmore than just a scaffoldrdquo FEBS Letters vol 581no 2 pp 175ndash179 2007

[136] J Moscat and M T Diaz-Meco ldquop62 at the crossroads of auto-phagy apoptosis and cancerrdquo Cell vol 137 no 6 pp 1001ndash10042009

[137] F Bardag-Gorce T Francis L Nan et al ldquoModifications in P62occur due to proteasome inhibition in alcoholic liver diseaserdquoLife Sciences vol 77 no 20 pp 2594ndash2602 2005

[138] K Nakaso Y Yoshimoto T Nakano et al ldquoTranscriptional acti-vation of p62A170ZIP during the formation of the aggregatespossible mechanisms and the role in Lewy body formation inParkinsonrsquos diseaserdquo Brain Research vol 1012 no 1-2 pp 42ndash512004

[139] E-L Eskelinen ldquoMaturation of autophagic vacuoles in mam-malian cellsrdquo Autophagy vol 1 no 1 pp 1ndash10 2005

[140] C M Hartford and M J Ratain ldquoRapamycin somethingold something new sometimes borrowed and now renewedrdquoClinical Pharmacology and Therapeutics vol 82 no 4 pp 381ndash388 2007

[141] C Takatsuka Y Inoue KMatsuoka and YMoriyasu ldquo3-Meth-yladenine inhibits autophagy in tobacco culture cells undersucrose starvation conditionsrdquo Plant and Cell Physiology vol45 no 3 pp 265ndash274 2004

[142] Y-TWuH-L TanG Shui et al ldquoDual role of 3-methyladeninein modulation of autophagy via different temporal patterns ofinhibition on class I and III phosphoinositide 3-kinaserdquo Journalof Biological Chemistry vol 285 no 14 pp 10850ndash10861 2010

[143] M F Ethier and J M Madison ldquoLY294002 but not wort-mannin increases intracellular calcium and inhibits calciumtransients in bovine and human airway smooth muscle cellsrdquoCell Calcium vol 32 no 1 pp 31ndash38 2002

Submit your manuscripts athttpwwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


main types according to the different pathway by which cargois delivered to the lysosome or vacuole chaperone-mediatedautophagy (CMA) microautophagy and macroautophagy[50]The last type can be divided into six principal steps initi-ation nucleation elongation closure maturation and degra-dation or extrusion [49] Through these steps cytoplasmicmaterials such as protein aggregates and organelles aresequestered by the phagophore a preautophagosomal mem-brane structure which thereafter expands and encloses itscargo to form a double-membrane vesicle the autophago-some [51] By its fusion with a lysosome it forms an autoly-sosome in which the enclosed cargo is degraded by acidhydrolases into biologically active monomers such as aminoacids that are subsequently reused to maintain cellular meta-bolic turnover and homeostasis [51]

Autophagy is involved in normal and diseased liver [52ndash55] Studies on autophagy in liver tissue following warmcold I-R remain controversial Some show defective andordecreased autophagy [27ndash30 32 34ndash37 39ndash41 45 47] in oth-ers hepatocellular autophagy is increased [26 31 33 38 42ndash44 46 48] Whether autophagy protects from or promotesliver injury following warm andor cold I-R remains to beelucidated

2 Pathomechanisms of Warm andor ColdLiver I-R Injury

Warm ischemia of the liver is due to oxygen deficiency causedby vascular occlusion of the liver during liver resection orhemorrhagic shock [2 10] Cold ischemia is observed duringliver transplantation in which the graft is subjected to warmand cold ischemia followed by a warm reperfusion phase thewarmcold ischemia sequence is due to vascular occlusion ofthe liver graft during its procurement from the donor andto graft storage in cold preservation solutions before livertransplantation [2 10] Graft implantation into the recipientrepresents the warm reperfusion phase all these phases leadto I-R injury of the liver [2 10] Pathomechanisms associatedwith warm I-R seem to differ from those after cold I-R in livertransplantation [57] Unlike warm liver I-R which resultsin primary damage to the hepatocytes cold I-R is mainlycharacterized by injury to the sinusoidal lining cells [58ndash60]

The introduction of oxygenated blood during reperfusionaggravates the ischemic insult which itself is mainly charac-terized by cellular adenosine triphosphate (ATP) depletionand results in perturbation of the cellular energy-dependentmetabolic and transport processes [1 61] Although the graftmetabolism is reduced during hypothermia with a prolongedtime period in which the anoxic cells can retain essentialmetabolic functions hypothermia may induce liver injury bydysfunction of the NaK ATPase membrane pump [61 62]The resulting intracellular influx and accumulation of sodiumand chloride lead to a perturbation of calcium homeostasisand to cell swelling [61 62] The inflammatory response toliver I-R involves neutrophils cytokines chemokines com-plement monocytes and macrophages [63ndash67] The reper-fusion process consists of two phases in the initial phaseactivated resident macrophages of the liver the Kupffer cells

induce oxidative stress mainly by reactive oxygen species(ROS) generation and in the later phase 6ndash24 hours follow-ing reperfusion recruited neutrophils release inflammatorymediators which can cause direct tissue damage [1] Kupffercells play a central role in the pathophysiologicalmechanismsof liver I-R Activated Kupffer cells release both ROS andcytokines including tumor necrosis factor alpha (TNF120572)Interleukin-1 (IL-1) and Interleukin-6 (IL-6) leading to gran-ulocytes accumulation in the sinusoidal space and causingmicrocirculatory disturbances [68] Accumulation of acti-vated neutrophils through release of oxidants and proteasesleads to hepatocyte damage ROS stimulate endothelial cellsto secrete platelet activating factor (PAF) [69] Monocytesand Kupffer cells-derived ROS activate redox-sensitive tran-scription factors AP-1 and nuclear factor kappa-B (NF-120581B) inendothelial cells and hepatocytes [65] Complement activa-tion products activate Kupffer cells during the initial phaseof liver injury and contribute to tissue inflammation as amembrane-attacking complex that stimulates the productionof proinflammatory cytokines and chemotactic agents whichoccur immediately after reperfusion [63] Complement canalso regulate the adaptative immunity [70 71] In fact theinflammation occurring during liver reperfusion is predomi-nantly an innate-immune-dominated response which mightinduce I-R injury in both parenchymal and nonparenchymalcells in situ and in liver transplants [66 67 72 73] Howeverthe cold preservation injury of the graft causes a likewisestrong adaptative immune response characterized by an earlyand massive T-cell influx into the ischemic liver graft [73ndash75]The conventional T lymphocytes CD4+ cells accumulatein the liver within 1 hour after reperfusion preceding anyneutrophil accumulation [76] CD4+ cells are activated byvarious Kupffer cell-derived products and lead to hepatocytesand sinusoidal endothelial cells damage [76] and finally livercells necrosis [22 23 77] Apoptosis seems to be a relevantdeath mechanism during warmcold liver I-R injury too[2 24 25] even if the studies are controversial on this issue[21 24 25] Recently the role of liver autophagy followingwarm andor cold liver I-R has been highlighted [78]

3 Autophagy and Warm andor ColdLiver I-R Injury

Beside necrosis [21ndash23] there are other modes of cell deathas apoptosis [2 24 25] and autophagy that may occur simul-taneously and or sequentially [26ndash48 78] following warmandor cold liver I-RWhile some studies have shown a defec-tive autophagy in hepatocytes following anoxiareoxygen-ation and in liver tissue following I-R [27ndash30 32 34ndash37 39ndash41 45 47] in others an increase of autophagy has beenobserved [26 31 33 38 42ndash44 46 48] (Table 1) with differenteffects on warm andor cold liver I-R injury (Figure 1)

31 Protective Role of Autophagy against Warm andorCold Liver I-R Injury

311 In Vitro and In Vivo Animal Studies The mammalianorthologue of yeast autophagy-related gene 6 (Atg6) Beclin 1


Table1Au

toph

agyandwarm

andor

cold

liver

I-Rinjuryinvitro

andin

vivo

anim

alandhu

man

studies

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Partialw

arm

60min

Decreased

(a)H

emin

+Ch

loroqu

ine

(b)H

emin

+Wortm

annin

Detrim

ental

Detrim

ental

Yunetal

2014

[47]

Mice

No

Partialw

arm

60min

Increased

Melaton

inProtectiv

eKa

ngetal

2014

[48]

Mice

No

Partialw

arm

45min

Increased

Ethylpyruvate

Protectiv

eShen

etal

2013

[46]

Mice

No

Totalw

arm

45min

Decreased

Carbam

azepine

Protectiv

eKim

etal

2013

[45]

Rat

No

Partialw

arm

60and

90min

Increased

Chloroqu

ine

Protectiv


ase)

Detrim

ental

(latereperfusionph

ase)

Fang

etal

2013

[44]

Rat

No

Partialw

arm

90min

Increased

Chloroqu

ine

Chloroqu

ine+

NAC

Detrim

ental

Protectiv

eSunetal

2013

[43]

Rat

No

Partialw

arm

60min

Increased

Lithium

Protectiv

eLiuetal

2013

[42]

Rat

Steatosis

Coldisc

hemia24

hDecreased

Mela

tonin+Trim

etazidine

Protectiv

eZa

oualietal

2013

[41]

Rat

Steatosis

Coldisc

hemia16h30

min

warm

reperfusionin

situ

Decreased

Detrim

ental

Gracia-Sancho

etal2013[40]

Hum

anNo

Hypoxiareoxygenatio

nof

hepatocytes

Decreased

3-MA

Protectiv

eBh

ogaletal

2012

[39]

Mice

No

(a)A

noxiareoxygenation

ofhepatocytes

Increased

Rapamycin

Protectiv

eWangetal

2012

[38]

(b)P

artia

lwarm

90min

Increased

Rapamycin

Protectiv

eCa

lcium

Calm

odulin-

kinase

IVKO

mice

No

Partialw

arm

60min

Decreased

Rapamycin

Protectiv

eEv

ankovich

etal2012[37]

Hum

anNo

Livertransplantatio

nDecreased

Ischem

icprecon

ditio

ning

Protectiv

eDegliEspo

stietal2011[35]

Old

mice

No

Totalw

arm

20min

Decreased

(a)A

LLM

Calpain2inhibitor

(b)A

deno

viraloverexpression

ofAtg4BandBe

clin1

Protectiv

eWangetal

2011[34]

Pig

No

Livertransplantatio

nDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2011[36]

Rat

Steatosis

Coldisc

hemia20

hDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2009

[30]

Mice

No

Partialw

arm

60min

Decreased

Cisplatin

Protectiv

eCardinaletal

2009

[32]


Table1Con

tinued

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Livertransplantatio

nIncreased

(a)W

ortm

annin

(b)L

Y294002

Protectiv

eGotoh

etal

2009

[33]

Hum

anPo

stchemotherapy

steatosis

TotalW

arm

Decreased

Ischem

icprecon

ditio

ning

Protectiv

eDom

artetal

2009

[29]

Old

mice

No

Partialw

arm

3060and

90min

Decreased

Rosig

litazon

eProtectiv

eSh

inetal

2008

[28]

Rat

No

(a)T

otalwarm

45min

Decreased

Protectiv

eKim

etal

2008

[27]

(b)A

noxiareoxygenation

ofhepatocytes

Decreased

(a)N

utrie

ntdepletion

(b)A

deno

viraloverexpression

ofAtg7

andBe

clin1

(c)A

LLM

Calpain

2inhibitor

Rat

No

Partialw

arm

120m

inIncreased

Protectiv

eCu

rsio

etal

2010

[31]

Rat

No

Livertransplantatio

nIncreased

Detrim

ental

Luetal

2005

[26]



Decreased autophagy

Increased autophagy










activation















darr ATP


darr AMPKuarr mTOR

uarr HMGB1

- Chloroquine



























































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table1Au

toph

agyandwarm

andor

cold

liver

I-Rinjuryinvitro

andin

vivo

anim

alandhu

man

studies

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Partialw

arm

60min

Decreased

(a)H

emin

+Ch

loroqu

ine

(b)H

emin

+Wortm

annin

Detrim

ental

Detrim

ental

Yunetal

2014

[47]

Mice

No

Partialw

arm

60min

Increased

Melaton

inProtectiv

eKa

ngetal

2014

[48]

Mice

No

Partialw

arm

45min

Increased

Ethylpyruvate

Protectiv

eShen

etal

2013

[46]

Mice

No

Totalw

arm

45min

Decreased

Carbam

azepine

Protectiv

eKim

etal

2013

[45]

Rat

No

Partialw

arm

60and

90min

Increased

Chloroqu

ine

Protectiv


ase)

Detrim

ental

(latereperfusionph

ase)

Fang

etal

2013

[44]

Rat

No

Partialw

arm

90min

Increased

Chloroqu

ine

Chloroqu

ine+

NAC

Detrim

ental

Protectiv

eSunetal

2013

[43]

Rat

No

Partialw

arm

60min

Increased

Lithium

Protectiv

eLiuetal

2013

[42]

Rat

Steatosis

Coldisc

hemia24

hDecreased

Mela

tonin+Trim

etazidine

Protectiv

eZa

oualietal

2013

[41]

Rat

Steatosis

Coldisc

hemia16h30

min

warm

reperfusionin

situ

Decreased

Detrim

ental

Gracia-Sancho

etal2013[40]

Hum

anNo

Hypoxiareoxygenatio

nof

hepatocytes

Decreased

3-MA

Protectiv

eBh

ogaletal

2012

[39]

Mice

No

(a)A

noxiareoxygenation

ofhepatocytes

Increased

Rapamycin

Protectiv

eWangetal

2012

[38]

(b)P

artia

lwarm

90min

Increased

Rapamycin

Protectiv

eCa

lcium

Calm

odulin-

kinase

IVKO

mice

No

Partialw

arm

60min

Decreased

Rapamycin

Protectiv

eEv

ankovich

etal2012[37]

Hum

anNo

Livertransplantatio

nDecreased

Ischem

icprecon

ditio

ning

Protectiv

eDegliEspo

stietal2011[35]

Old

mice

No

Totalw

arm

20min

Decreased

(a)A

LLM

Calpain2inhibitor

(b)A

deno

viraloverexpression

ofAtg4BandBe

clin1

Protectiv

eWangetal

2011[34]

Pig

No

Livertransplantatio

nDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2011[36]

Rat

Steatosis

Coldisc

hemia20

hDecreased

Hypotherm

icrecond

ition

ing

bygaseou

soxygen

Protectiv

eMinor

etal

2009

[30]

Mice

No

Partialw

arm

60min

Decreased

Cisplatin

Protectiv

eCardinaletal

2009

[32]


Table1Con

tinued

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Livertransplantatio

nIncreased

(a)W

ortm

annin

(b)L

Y294002

Protectiv

eGotoh

etal

2009

[33]

Hum

anPo

stchemotherapy

steatosis

TotalW

arm

Decreased

Ischem

icprecon

ditio

ning

Protectiv

eDom

artetal

2009

[29]

Old

mice

No

Partialw

arm

3060and

90min

Decreased

Rosig

litazon

eProtectiv

eSh

inetal

2008

[28]

Rat

No

(a)T

otalwarm

45min

Decreased

Protectiv

eKim

etal

2008

[27]

(b)A

noxiareoxygenation

ofhepatocytes

Decreased

(a)N

utrie

ntdepletion

(b)A

deno

viraloverexpression

ofAtg7

andBe

clin1

(c)A

LLM

Calpain

2inhibitor

Rat

No

Partialw

arm

120m

inIncreased

Protectiv

eCu

rsio

etal

2010

[31]

Rat

No

Livertransplantatio

nIncreased

Detrim

ental

Luetal

2005

[26]



Decreased autophagy

Increased autophagy










activation















darr ATP


darr AMPKuarr mTOR

uarr HMGB1

- Chloroquine



























































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table1Con

tinued

Species

Preexisting

liver

disease

Experim

entalliver

I-R

mod

elEff

ectson

autoph

agy

Mod

ulationof

autoph

agy

byinhibitors

Mod

ulationof

autoph

agyby

stim

ulators

Effectson

I-Rinjury

Authorsa

ndreferences

Rat

No

Livertransplantatio

nIncreased

(a)W

ortm

annin

(b)L

Y294002

Protectiv

eGotoh

etal

2009

[33]

Hum

anPo

stchemotherapy

steatosis

TotalW

arm

Decreased

Ischem

icprecon

ditio

ning

Protectiv

eDom

artetal

2009

[29]

Old

mice

No

Partialw

arm

3060and

90min

Decreased

Rosig

litazon

eProtectiv

eSh

inetal

2008

[28]

Rat

No

(a)T

otalwarm

45min

Decreased

Protectiv

eKim

etal

2008

[27]

(b)A

noxiareoxygenation

ofhepatocytes

Decreased

(a)N

utrie

ntdepletion

(b)A

deno

viraloverexpression

ofAtg7

andBe

clin1

(c)A

LLM

Calpain

2inhibitor

Rat

No

Partialw

arm

120m

inIncreased

Protectiv

eCu

rsio

etal

2010

[31]

Rat

No

Livertransplantatio

nIncreased

Detrim

ental

Luetal

2005

[26]



Decreased autophagy

Increased autophagy










activation















darr ATP


darr AMPKuarr mTOR

uarr HMGB1

- Chloroquine



























































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of


















Decreased autophagy

Increased autophagy










activation















darr ATP


darr AMPKuarr mTOR

uarr HMGB1

- Chloroquine



























































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






































































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

























































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of














































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



































apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

























apoptosis

Kim et al2013 [45]


Fang et al2013 [44] (+)



necrosis by NAC

Liu et al2013 [42]









Wang et al2011 [34] (+)



ATP increase



Gotoh et al2009 [33] (+)





Lu et al2005 [26] (+)






2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















2S asso-











4 Conclusion








References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of






















References



























































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















































































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















































































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















































































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Documents

Review Article Autophagy and Liver Ischemia-Reperfusion Injurydownloads.hindawi.com/journals/bmri/2015/417590.pdf · 2019-07-31 · In Vitro and In Vivo Animal Studies. e mammalian