Phytomedical Effect of Soy Protein and Soy Isoflavone on Hypercholesterolemia and-Or Atherosclerosis-24_057

7/31/2019 Phytomedical Effect of Soy Protein and Soy Isoflavone on Hypercholesterolemia and-Or Atherosclerosis-24_057

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*1 1 1

Aram Joeng1

2, 3

Rosario M. Perez 3 Romualdo L. Carrasco 3 2, 3

1 2 3INA Research Philippines

Phytomedical Effect of Soy Protein and Soy Isoflavoneon Hypercholesterolemia and/or Atheroscelerosis :

Its Molecular Pathophysiological Mechanism in Monkeys

Shin NAKAMURA1, Fusako MITSUNAGA 1, Maki OKADA1,Aram JOENG 1, Takashi HAYASHI 2, 3, Rosario M. PEREZ 3, Romualdo L. CARRASCO3,

and Hiroshi NAKAGAWA 2, 3.

1Primate Research Institute, Kyoto University, Inuyama 484-85062Ina Research Inc., Nagano 399-4501 3INA Research Philippines, Philippines.

ABSTRACT

Soy protein ( -Conglycinine) and soy isoflavone (Soyaflavone HG) are known toregulate hypercholesterolemia and its mediated atherosclerosis. In the current studywe examined the phytomedical effect of -Conglycinine and Soyaflavone using anestablished monkey hypercholesterolemia/atherosclerosis model, in normal male,female and ovariectomized (OVX)-female monkeys. The molecular mechanism of thephyto-medicial function of the soy products was investigated by analyses of blood lipidmarkers (total Chol, LDL-C, HDL-C, GOT, GPT, Glu, CPK, TG, Apo-Al, Apo-B, and

Apo-E). To elucidate the genetic mechanism, gene expression profiling analyses of aninflammatory and thrombotic gene (TF) and lipid metabolic genes (LDL-R, VLDL-R,HMG-CoA, SR) were performed by RT-PCR. -Conglycinine and Soyaflavone bothsuppressed upregulated TF gene expression in hypercholesterolemia, suggesting a newmechanism of these soy products in hypercholesterolemia/atherosclerosis. Soy Protein

Research, Japan 6 , 57-62, 2003.

Key words : -conglycinine, soyaflavone, hypercholesterolemia, monkey model, geneexpression profiling

484-8506 41-2


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-

1, 2)

-conglycinineSoyaflavone HG

OVX-Table 1

AS

400700 mg/mL 22AS

PSSoyaflavone HG

-conglycinine

100 mg/head/daySoyaflavone HG 7.5 g/head/day -

conglycinine

WBC RBC HGBHCT MCV MCH Plt Ret WBC differential countSt Seg Eo Ba Ly Mo) PT APTT

HDL-C LDL-CGOT GPT Glu CPK TG Apo-Al

Apo-B Apo-E 3)

LDL/HDL ratio 3.0hypercholesterolemia

PBS Ficol/Conray1.077 PBMC;monocytes/lymphocytes PBMC

PBMC Isogen total RNAIsogen DNA

QIAGEN-Kit RNADNA DNA- total RNA

RT-PCRRT 100 ng DNA- total RNAOligo-dT primer cDNAcDNA template GAPDH TFLDL-R HMG SR VLDL-R DNA fragment

PCR primer PCRFig. 1 PCR PAGE-

DNA SYBR Green1

106PCR

AS400700 mg/100 mL

Table 1. Composition of the special diet AS to inducehypercholesterolemia in monkey model

Crude protein 21.1%Crude fat 16.7%Calories 418 kcal

Powder ingredient (AS base) 45.1%Salt-free butter 15.0%Sucrose 25.0%Milk casein 10.0%Cholesterol 0.3%Minerals 3.5%

Vitamins 1.0%(A, D3, E, K, B complex)Vitamin C 0.1


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LDL/HDL 3.0Tale 2

AS PSLDL/HDLOVX- OVX-

LDL/HDL Fig. 2

7.5 g/head/day

-conglycinine OVX-100 mg/head/day Soyaflavone

HG

Fig.1. Gene expression analyses by RT-PCR.

Table 2. Feeding conditions and profiles of blood cholesterol level in monkey

Conditions

1104(IM0002)

070

210210

52.552.552.5

000

52.500

52.552.552.521212152.552.552.5

PretestPretestWeek 1Week 2Week 3Week 4Week 5Week 6Week 7Week 8Week 9Week 10Week 12Week 14Week 15Week 16Week 17Week 18Week 19Week 20Week 21Week 22

7.056.857.207.407.457.307.207.157.107.056.957.106.857.107.057.157.057.207.207.257.257.00

167224378595786832914806522429538473361543548607586548459448631631

100169266351371408360233192240211161242245271262245205200282282

7985

10241844051342628533439595545512625367044

70778739261819192331382629332628302331273123

78110262495668795754666417338421397278402419475464439405377515512

97147291556760814895787499398500447332510522579143525428421600608

2.42.94.3

15.330.246.248.142.422.713.814.218.212.416.521.121.7

5.823.814.816.620.427.4

1.11.43.0

12.725.744.239.735.118.110.911.115.3

9.612.216.117.015.519.113.114.016.622.3

1105(IM0229)

070

210210

52.5210210210

00

105210105157.5157.5157.5

10552.552.552.552.552.5

PretestPretestWeek 1Week 2Week 3Week 4Week 5Week 6Week 7Week 8Week 9Week 10Week 12Week 14Week 15Week 16

Week 17Week 18Week 19Week 20Week 21Week 22

7.357.157.157.107.257.157.157.107.157.157.257.357.157.156.906.85

6.806.856.956.856.756.70

120131395334505523628750422244199275245480538633

675687621544651706

100302255385399479573322186152210187366411483

515524474415497539

1714511525131514

810121418282421

262724163327

44565145483836302328413735363638

343035272928

6062

254226368444489567321184138205180342369439

475492448381470477

7675

344289457485592720399216158238210444502595

641657586517622678

2.71.77.77.4

10.513.817.425.018.3

8.74.97.47.0

13.314.916.7

19.922.917.720.122.425.2

1.41.15.05.07.7

11.713.618.914.0

6.63.45.55.19.5

10.311.6

14.016.412.814.116.217.0

Animal

Number

AS Given

(g)

Time Point Body weight

(kg)

CHOL

mg/100 mL

CHOLage value

%

TG

mg/100 mL

HDL-C

mg/100 mL

LDL-C

mg/100 mL

Non-HDL-C

mg/100 mL

C/H

ratio

LDL/HDL

ratio


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HDL-C LDL-CFig. 3 LDL/HDL

-conglycinineLDL/HDL 48

LDL/HDL 3.0Soyaflavone HG

OVX- LDL/HDL

TG Apo-Al Apo-B Apo-E

-conglycinine SoyaflavoneHG LDL/HDL

OVXSoyaflavone HG

Fig. 1tissue factor TF

)4) 3-hydroxy-3-methylgulutaryl coenzyme A reductase (HMG) 5) lowdensity lipoprotein receptor (LDL-R) 6) very lowdensity lipoprotein receptor (VLDL-R) 7)

scavenger receptor (SR) 7)

PBMC /

Pre CholChol + SP (soy protein)

RT-PCR Fig. 4A Fig. 4B -conglycinine

Fig. 4A Soyaflavone HGFig. 4B

-conglycinine TFOVX-

Soyaflavone HG TF

LDL-R VLDL-R SR

Fig. 2. Comparsion of hypercholesterolemia responseamong OVX-female, normal female and normalmale monkeys.

Fig. 3. Effect of oral administration of soy product on blood cholesterol level in OVX-female and normalmale monkeys fed the high cholesterol diet.

OVX-female monkeys were orally given with 100mg/head/day of Soyaflavone suspension andnormal male were orally given 7.5 g/head/day of -conglycinine suspension for 8 weeks.

OVX-

TF -conglycinine Soyaflavone

HGTF

VII

8, 9) -conglycinine Soyaflavone HGTF

TF

PBMC


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OVX-

Fig. 4A. Effect of a soy protein, -conglycinine (Con),on gene expression in PBMC of malehypercholesterolemia monkey. Total RNA wasisolated from PBMC separated from 5 mLmonkey whole blood. RT-PCR reaction anddetection of PCR product were performedunder the conditions as mentioned in Fig. 1.Relative gene expression of each gene wasexpressed by densitometry.

Fig. 4B. Effect of a soy isoflavone, Soyaflavone (IF), ongene expression in PBMC of OVX-femalehypercholesterolemia monkey. Total RNA wasisolated from PBMC separated from 5 mL monkeywhole blood. RT-PCR reaction and detection of PCR product were performed under the conditionsas mentioned in Fig. 1. Relative gene expression of each gene was expressed by densitometry.

AS

-conglycinineSoyaflavone HG

-conglycinineSoyaflavone HGLDL-C HDL-C GOT GPT Glu CPK TG Apo-Al Apo-B Apo-E

PBMC TF HMG-CoA LDL-R VLDL-R SR TF

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Phytomedical Effect of Soy Protein and Soy Isoflavone on Hypercholesterolemia and-Or Atherosclerosis-24_057