Dose-dependent Decaffeinated Green Tea Extract Administration

Improved Hyperglycemia through Modulation of IRS-1 and GLUT-4

Genes Expression in Metabolic Syndrome Rat Model

Dwi Adi Nugroho

, Mifetika Lukitasari

Marlita Marlita

Mohammad Saifur Rohman

Nashi Widodo

, Inggita Kusumastuty

and Nur Ida Panca Nugrahini

Department of Herbal Medicine, Cardiovascular research group, Faculty of Medicine, Brawijaya University, Malang,

Indonesia.

Department of Nursing, Faculty of Medicine, Brawijaya University,Malang, Indonesia.

Department of cell culture, Animal Physiology, Structure and Development Laboratory, Faculty of Mathematics and

Natural Science, Brawijaya University, Malang, Indonesia.

Department of Cardiology and Vascular Medicine, Faculty of Medicine, Brawijaya University-Saiful Anwar General

Hospitalt, Malang, Indonesia.

Department of Biology, Faculty of Mathematics and Natural Science, Brawijaya University,, Malang, Indonesia.

Department of Nutrition, Faculty of Medicine, Brawijaya University, Malang, Indonesia.

Departement Agricultural Product Technology, Brawijaya University, Malang, Indonesia.

Keywords: Green tea, Hyperglycemia, IRS-1, GLUT-4, Metabolic Syndrome

Abstract: Hyperglycemia is a major disorder in metabolic syndrome. Skeletal IRS-1 dan GLUT-4 expression are the

key target in hyperglycemia improvement. This study aimed to investigate green tea extract's effect on

hyperglycemia improvement in metabolic syndrome rat models. Twenty Sprague Dawley Metabolic

Syndrome Rat Model weighed 300 – 400 grams were divided into GTE 200 (n=5) and GTE 400 (n=5) groups.

Moreover, as control groups, ten rats were divided into normal control (NC) (n=5) and metabolic syndrome

(MS) (n=5) groups. Rats in the GTE 200 and 400 groups were treated once daily with green tea extract at a

dose of 200 and 400 mg/bw.t, respectively. The extract was administered for 9 weeks through oral gavage.

RT-PCR methods analyzed skeletal IRS-1 and GLUT-4 gene expression. This study showed that the fasting

blood glucose of GTE 200 dan GTE 400 was significantly lower than those of the MS group (p<0.001 and

p<0.001, respectively). In addition, GTE 400 group had the lowest fasting blood glucose. Moreover, skeletal

IRS-1 dan GLUT-4 gene expression was significantly higher in the GTE 200 and GTE 400 group than those

of the MS group. In contrast, the GTE 400 group's gene expression was the highest among all groups (p<0.000

and p<0.002, respectively). Administration of green tea extract improved hyperglycemia in a metabolic

syndrome rat model in a dose-dependent manner through skeletal IRS-1 dan GLUT-4 gene expression

modulation.

https://orcid.org/0000- 0002-6195-9771

https://orcid.org/0000- 0002-3971-7418

https://orcid.org/0000- 0001-7955-1320

https://orcid.org/0000- 0001-6461-2223

https://orcid.org/0000- 0002-1126-498X

https://orcid.org/0000- 0002-0481-4541

https://orcid.org/0000- 0003-2781-2554

Nugroho, D., Lukitasari, M., Marlita, M., Rohman, M., Widodo, N., Kusumastuty, I. and Panca Nugrahini, N.

Dose-dependent Decaffeinated Green Tea Extract Administration Improved Hyperglycemia through Modulation of IRS-1 and GLUT-4 Genes Expression in Metabolic Syndrome Rat Model.

DOI: 10.5220/0010487900690074

In Proceedings of the 1st Jenderal Soedirman International Medical Conference in conjunction with the 5th Annual Scientiﬁc Meeting (Temilnas) Consortium of Biomedical Science Indonesia

(JIMC 2020), pages 69-74

ISBN: 978-989-758-499-2

1 INTRODUCTION

Metabolic syndrome (MS) is a complex multifactorial

disorder that increases the risk of cardiovascular

disease and diabetes mellitus type 2. MS prevalence

ranges between 10% and 84%, depending on the age,

gender, race, ethnicity, and MS criteria.(Alberti et al.,

2009) The World Health Organization, International

Diabetes Federation, and National Cholesterol of

Adult Treatment Panel III (NCEP-ATP III) have

determined specific criteria for MS, which includes

central obesity, high blood pressure, high triglyceride

(TG) levels, low high-density lipoprotein (HDL), and

high glucose levels.(Grundy, 2016) There is no single

treatment for MS, and natural products have gained

attention as potential treatments have

increased.(Cerezo et al., 2013; Rohman, 2011)

Tea, the most widely consumed beverage

globally, has attracted significant public interest for

its potential health benefits.(Cheng et al., 2020;

Department of Biology, College of Science,

University of Baghdad, Baghdad-Iraq & Al-Hilfy,

2012).

Green tea contains caffeine and polyphenolic

compounds, known as catechins. The most abundant

catechin found in green tea is (−)-epigallocatechin-3-

gallate (EGCG). Tea catechins are also thought to be

useful for their antiobesity, antioxidant,

antihypertensive, anticarcinogenic, and

hypocholesterolemic action. Several studies have

described the beneficial effects of tea constituents in

animal models of MS.(Ding et al., 2017; Gan et al.,

2015; Riegsecker et al., 2013).

Moreover, the skeletal insulin receptor substrate

(IRS)/glucose transporter-4 (GLUT-4) pathway has

shown to improve hyperglycemia in metabolic

disorders in animal models.(Casanova et al., 2019;

Cheng et al., 2020) A previous study reported that

oral administration of green tea extract (GTE)

significantly improved hyperglycemia and increased

insulin sensitivity in patients with metabolic

disorders.(Wu et al., 2004)

Previous study by Cao et al suggested that green

tea extract administration with the dose of 1-2 g/body

weight increased IRS1 and GLUT4 mRNA level in

skeletal tissue of high fructose diet rats. Moreover,

Wu et al suggested that intravenous administration of

0,5 g/100 ml green tea extract alleviated

hyperglycemia and increased GLUT mRNA in high

fat diet induced rat model for 12 weeks. However, as

our knowledge there were limited data regarding the

effect of green tea administration on metabolic

syndrome rat model.

Therefore, this study aimed to investigate the

effect of decaffeinated green tea extract in high fat,

high sucrose diet, and low dose streptozococin

induced metabolic syndrome rat model for 9 weeks.

We hypothesized that decaffeinated GTE modulates

IRS/GLUT-4 gene expression and improves

hyperglycemia in MS rats.

2 MATERIALS AND METHODS

2.1 Extraction of Green Tea

Green tea was extracted from the young leaves of

green tea. Green tea leaves were sorted to obtain high-

quality seeds. A dryer cabinet set at 50°C was used to

dry 500 g of green tea leaves for 8 h to obtain

simplicia with 8%–10% water content. The simplicia

was mashed with a blender and macerated with

methanol to produce a crude extract. The crude

extract was then filtered using a filter cloth to separate

the liquid from the solid phase. The liquid phase was

concentrated using a rotary evaporator at a

temperature of ±40°C. The concentrated liquid phase

was partitioned using butanol, water, and

acetilacetate Finally, column chromatography was

performed using silica gel as the static phase, and the

filtered product was evaporated.(Banerjee &

Chatterjee, 2014).

2.2 Animal Care and Experimental

Protocol

Twenty male Sprague Dawley rats were purchased

from the National Agency of Drug and Food Control

of Indonesia. They were housed in standard cages and

placed in a room where temperature was maintained

at 25°C ± 1°C and relative humidity at 50% ± 1%,

with a 12 h light/dark cycle. During a 1-week

acclimatization period, all rats consumed a normal

pellet diet and tap water ad libitum. The rats then

received a high sucrose, fat, and sodium diet for 9

weeks and an intraperitoneal streptozotocin injection

(30 mg/body weight [BW]) in the second and third

weeks. Rats with >126 mg/dL blood glucose, >150

mg/dL triglyceride, high systolic blood pressure

(≥140 mm/Hg), and reduced HDL levels (<40 mg/dL)

were confirmed as MS rats based on NCEP-ATP III

criteria.(Saifur Rohman et al., 2017) The rats were

divided into four weight-matched groups (𝑛 = 5): the

normal control (NC), Metabolic syndrome (MS),

metabolic syndrome with 200 mg/b.wt GTE (GTE

200), and metabolic syndrome with 400 mg/bw.t GTE

(GTE 400). The extract was given via oral gavage

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

daily. Extract dose was given in milliliters based on

the weekly BW measurement. Food and water intake

were recorded daily. At the end of the experimental

period, animals were anesthetized with ether

following a 12 h fasting period. Blood samples were

drawn from the heart into a micro-centrifuge tube,

and serum samples were obtained by centrifugation at

4000 × g for 15 min at 4°C. The protocol was

reviewed and approved by the ethics committee of the

Faculty of Medicine, Brawijaya University.

2.3 Physiological Measurement

Daily food and fluid intake were measured on

everyday basis, and BW was measured weekly. Food

and fluid intake of each rat was measured by

subtracting the amount initially provided by the

remaining amount in the cage.

2.4 Biochemical Analysis

The serum concentrations of fasting blood glucose,

TG, and HDL cholesterol were measured

enzymatically using commercial kits (Biolabo,

France).

2.5 Blood Pressure Measurements

Blood pressure was measured using the tail-cuff

method with a sphygmomanometer at baseline and at

the end of the experiment. Three readings were taken

consecutively, which were averaged to provide a final

systolic Blood Pressure (SBP) reading.

2.6 Gene Expression Analysis

Total RNA was extracted from skeletal tissues with

TRIzol Reagent (Invitrogen, USA). The total RNA (2

g) was reverse transcribed using the SuperScript

First-Strand Synthesis System (Invitrogen, USA).

Primers were designed according to the sequences in

GenBank as follows: -actin F: “TAC AAC CTC

CTT GCA GCT CC,” R: “GGA TCT TCA TGA

GGT AGT CAG TC;” IRS-1 F: “AAG CAC CTG

GTG GCT CTC TA,” R: “TCA GGA TAA CCT

GCC AGA CC;” and GLUT-4 F: “CTT CCT TCT

ATT TGC CGT CCT C,” R: “GCT GCT GTT TCC

TTC ATC CTG.” Standard 25 L polymerase chain

reaction (PCR) with 2 L of the reverse transcriptase

was performed using the following parameters: 95°C,

40 s, annealing temperature, 40 s, 72°C, 45 s, for 27

cycles with TaKaRa Ex Taq Hot Start Version

(TaKaRa, Japan) in an MJ Research PTC-200 Peltier

Thermal Cycler. The PCR reaction product (10 L)

was separated using 2% agarose gels by

electrophoresis. Densitometric quantification of the

band intensities was conducted using NIH Image J

software.

2.7 Statistical Analysis

All data were analyzed using Statistical Package for

Social Sciences (version 22) and are presented as

mean ± standard deviation. The data were subjected

to a one-way analysis of variance, independent t-test,

and paired t-test and used a significance level of p <

0.05.

3 RESULTS

3.1 Baseline Characteristics

Before treatment, the MS rats were characterized by

obesity, high systolic blood pressure, high TG,

hyperglycemia, and low HDL cholesterol, as shown

in Table 1. These characteristics were similar to the

MS characteristics observed in humans, according to

the NCEP-ATP III criteria.

Table 1. Baseline Characteristics Metabolic Syndrome Rat

Model.

Experimental group

Paramater

NC MS p

Body Weight

295,80±5,11 366,2±7,59 0,012

Food Intake

19,40±0,66 22,6±1,67 0,000

Water Intake

26,85±1,92 45,4±14,02 0,001`

Blood Glucose

101,2±4,32 250±40,74 0,000

Triglyceride

81,20±6,76 252,8±66,83 0,000

HDL

44,00±3,46 31,40±6,87 0,000

Blood pressure

124,6±5,5 152,6±7,12 0,000

Values are mean ± SD, n = 5. data was analyzed by

dependent t-test.

NC : Normal Control;

MS : Metabolic syndrome Induces

3.2 Effects of GTE on Fasting Blood

Glucose Levels

The effect of GTE on fasting blood glucose levels in

the experimental animals is presented in Table 2. The

level of fasting blood glucose was not significantly

Dose-dependent Decaffeinated Green Tea Extract Administration Improved Hyperglycemia through Modulation of IRS-1 and GLUT-4

Genes Expression in Metabolic Syndrome Rat Model

different between any of the groups at baseline.

Following 9 weeks of intervention, fasting blood

glucose levels in all GTE groups significantly

decreased from baseline (p < 0.001). Furthermore, all

interventional groups showed greater decrease in

fasting blood glucose levels compared to that of the

MS group (p < 0.05). The GTE 400 group had the

lowest fasting blood glucose level.

Table 2. The comparison of fasting blood glucose among

groups.

Experimental

group

Fastin

Blood Glucose

Pre Post

NC 101,2±4,32 92,40±9,50

MS 250±40,74 283,2±31,92*

GTE 200 243,89±29,21 196,41±19,19*a

GTE 400 223,40±18,69 181,95±14,95*ab

Values are mean ± SD, n = 5. data was analyzed by

dependent t-test.

* : significant between pre test and postest ( p < 0,05)

a : significant compared to that MS group ( p < 0,05)

b : significant compared to that GTE 200 group ( p < 0,05)

NC : Normal Control;

MS : Metabolic syndrome Induces

EGCG 200 : Metabolic syndrome with green tea

extract

200 mg/kg.bw.t

EGCG 400 : Metabolic syndrome with green tea

extract

400 mg/kg.bw.t

3.3 Effect of GTE on IRS-1 and

GLUT-4 Gene Expression

We examined skeletal mRNA gene expression to

determine the effect of 9 weeks of decaffeinated GTE

administration on IRS-1 (Figure 1a) and GLUT-4

gene expression (Figure 1b). In skeletal tissue, IRS-1

and GLUT-4 gene expression were significantly

higher in all GTE groups compared with those of the

MS group (p < 0.001 and p < 0.002, respectively).

Moreover, gene expression in the GTE 400 group was

the highest among all groups.

4 DISCUSSION

We investigated the effect of decaffeinated GTE on

hyperglycemia by modulation of IRS-1 and GLUT-4

gene expression. A recent study was conducted in a

rat model that the criteria of MS (hyperglycemia,

elevated triglyceride level, decreased HDL level, and

hypertension) as presented in baseline characteristics

in Table 1.(Saifur Rohman et al., 2017) MS was

confirmed by high IRS-1 and GLUT-4 gene

expression and significant reduction in fasting blood

glucose levels in the GTE 200 and GTE 400 groups.

The development of metabolic syndrome rat model in

this study was different from previous study that used

high fructose diet or high fat diet only or genetically

modified rat model. In fact, the rat model in this study

represented the features of metabolic syndrome in

human, such as hyperglycemia, hypertension, and

dyslipidemia.

Hyperglycemia improved after 9 weeks of

decaffeinated GTE administration. Furthermore, we

(a) (b)

Figure 1. Effects of green tea extract on IRS-1 (a) and Glut-4 gene expression (b).

Values are mean ± SD, n = 5. data was analyzed by independent t-test.

* : significant compared to that MS group (p< 0,05)

# : significant compared to that GTE 200 group (p< 0,05)

NC : Normal Control;

MS : Metabolic syndrome Induces

EGCG 200 : Metabolic syndrome with green tea extract 200 mg/kg.bw.t

GCG

etabo

tea

act

g.bw.t

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )

revealed that one mechanism of hyperglycemia

improvement was via the skeletal IRS/GLUT-4

pathway.(Boucher et al., 2014; Chang et al., 2004)

Modulation of IRS-1 may have increased GLUT-4

translocation, which induced the reuptake of plasma

glucose and improved hyperglycemia. A study by

Jang showed that EGCG in green tea reduces fasting

glucose and increases insulin and GLUT-4 expression

levels in skeletal muscle and adipose tissue.(Fu et al.,

2017; Jang et al., 2013) Another study showed that

the administration of GTE regulated the expression of

genes involved in insulin-signaling pathways in the

muscle tissue of rats with MS induced by a high-

fructose diet.(Wu et al., 2004) GTE significantly

increased mRNA levels of IRS1 and GLUT4 in the

muscle tissue. An in vitro study by Zhang showed that

GTE-rich EGCG improved IRS-1 and GLUT-4 gene

expression in L6 muscle cells after dexamethasone

induction (Zhang et al., 2010).

A study by Cao showed that GTE at 1 or 2 g/kg

BW regulates IRS-1 and GLUT-4 gene expression in

rats that are fed a fructose-rich diet. Moreover,(Cao et

al., 2007) a study by Cheng et al. showed that

administration of 200 mg/b.wt green tea extract

decreases fasting glucose, enhances the expression

and translocation of GLUT-4, and activates IRS-1

through decreased pSer612IRS-1 expression.(Cheng

et al., 2020)

Hyperglycemia alleviation after green tea extract

administration might achieved through other pathway

such as adiponectin receptor-AMPK pathway,

inflammation inhibition pathway by inhibiting

gluconeogenesis factor such as FOX-O and PEPCK

in hepatic, skeletal, and adipocyte tissue.

Our study showed that GTE 400 had a larger

effect on hyperglycemia compared to that of GTE

200, demonstrating a dose effect.(Lukitasari et al.,

2018) We used decaffeinated grren tea extract

because caffeine may induce palpitations and

increase blood homocysteine, which reduced the

antioxidant effect of EGCG that was abundantly

obtained from tea. Therefore, decaffeinated GTE

might minimize these side effects.(Roberts et al.,

2015)

5 CONCLUSIONS

Our study revealed the beneficial effect of

decaffeinated GTE on hyperglycemia via the

modulation of IRS-1 and GLUT-4 receptor gene

expression in the MS rat model.

ACKNOWLEDGMENTS

Thanks to Cardiovascular Research Group, Medical

Faculty of Brawijaya University, Biology

Mathematics and Natural Sciences Faculty of

Brawijaya University, and the Ministry of Research,

Technology, and Higher Education of the Republic of

Indonesia.

REFERENCES

Alberti, K. G. M. M., Eckel, R. H., Grundy, S. M., Zimmet,

P. Z., Cleeman, J. I., Donato, K. A., Fruchart, J.-C.,

James, W. P. T., Loria, C. M., Smith, S. C.,

International Diabetes Federation Task Force on

Epidemiology and Prevention, Hational Heart, Lung,

and Blood Institute, American Heart Association,

World Heart Federation, International Atherosclerosis

Society, & International Association for the Study of

Obesity., 2009. Harmonizing the metabolic syndrome:

A joint interim statement of the International Diabetes

Federation Task Force on Epidemiology and

Prevention; National Heart, Lung, and Blood Institute;

American Heart Association; World Heart Federation;

International Atherosclerosis Society; and International

Association for the Study of Obesity. Circulation,

120(16), pp. 1640–1645.

Banerjee, S., & Chatterjee, J., 2014. Efficient extraction

strategies of tea (Camellia sinensis) biomolecules.

Journal of Food Science and Technology, 52(6), pp.

3158-68 ,

Boucher, J., Kleinridders, A., & Kahn, C. R. (2014). Insulin

Receptor Signaling in Normal and Insulin-Resistant

States. Cold Spring Harbor Perspectives in Biology,

6(1).

Cao, H., Hininger-Favier, I., Kelly, M. A., Benaraba, R.,

Dawson, H. D., Coves, S., Roussel, A. M., & Anderson,

R. A., 2007. Green Tea Polyphenol Extract Regulates

the Expression of Genes Involved in Glucose Uptake

and Insulin Signaling in Rats Fed a High Fructose Diet.

Journal of Agricultural and Food Chemistry, 55(15),

pp. 6372–6378.

Casanova, E., Salvadó, J., Crescenti, A., & Gibert-Ramos,

A., 2019. Epigallocatechin Gallate Modulates Muscle

Homeostasis in Type 2 Diabetes and Obesity by

Targeting Energetic and Redox Pathways: A Narrative

Review. International Journal of Molecular Sciences,

20(3).

Cerezo, C., Segura, J., Praga, M., & Ruilope, L. M., 2013.

Guidelines Updates in the Treatment of Obesity or

Metabolic Syndrome and Hypertension. Current

Hypertension Reports, 15(3), pp. 196–203.

Chang, L., Chiang, S.-H., & Saltiel, A. R., 2004. Insulin

Signaling and the Regulation of Glucose Transport.

Molecular Medicine, 10(7–12), pp. 65–71.

Cheng, J., Tan, Y., Zhou, J., Xiao, L., Johnson, M., & Qu,

X., 2020. Green tea polyphenols ameliorate metabolic

Dose-dependent Decaffeinated Green Tea Extract Administration Improved Hyperglycemia through Modulation of IRS-1 and GLUT-4

Genes Expression in Metabolic Syndrome Rat Model

abnormalities and insulin resistance by enhancing

insulin signalling in skeletal muscle of Zucker fatty rats.

Clinical Science, 134(10), pp. 1167–1180.

Department of Biology, College of Science, University of

Baghdad, Baghdad-Iraq, & Al-Hilfy, J. H. Y., 2012.

Effect of Green Tea Aqueous Extract on Body Weight,

Glucose Level, and Kidney Functions in Diabetic Male

Albino Rats. Journal of Al-Nahrain University Science,

15(3), pp. 161–166.

Ding, S., Jiang, J., Yu, P., Zhang, G., Zhang, G., & Liu, X.,

2017. Green tea polyphenol treatment attenuates

atherosclerosis in high-fat diet-fed apolipoprotein E-

knockout mice via alleviating dyslipidemia and up-

regulating autophagy. PLOS ONE, 12(8), e0181666.

Fu, Q.-Y., Li, Q.-S., Lin, X.-M., Qiao, R.-Y., Yang, R., Li,

X.-M., Dong, Z.-B., Xiang, L.-P., Zheng, X.-Q., Lu, J.-

L., Yuan, C.-B., Ye, J.-H., & Liang, Y.-R., 2017.

Antidiabetic Effects of Tea. Molecules (Basel,

Switzerland), 22(5).

Gan, L., Meng, Z., Xiong, R., Guo, J., Lu, X., Zheng, Z.,

Deng, Y., Luo, B., Zou, F., & Li, H., 2015. Green tea

polyphenol epigallocatechin-3-gallate ameliorates

insulin resistance in non-alcoholic fatty liver disease

mice. Acta Pharmacologica Sinica, 36(5), pp. 597–605.

Grundy, S. M., 2016. Metabolic syndrome update. Trends

in Cardiovascular Medicine, 26(4),pp. 364–373.

Jang, H.-J., Ridgeway, S. D., & Kim, J., 2013. Effects of

the green tea polyphenol epigallocatechin-3-gallate on

high-fat diet-induced insulin resistance and endothelial

dysfunction. American Journal of Physiology-

Endocrinology and Metabolism, 305(12), pp. E1444–

E1451.

Lukitasari, M., Nugroho, D. A., & Rohman, M. S., 2018.

Green Tea Extract Administration Had A Beneficial

Effect On Ppar Alpha And Ppar Gamma Gene

Expression In Metabolic Syndrome Rat Model: Journal

of Hypertension, 36, e9.

Riegsecker, S., Wiczynski, D., Kaplan, M. J., & Ahmed, S.,

2013. Potential Benefits of Green Tea Polyphenol

EGCG in the Prevention and Treatment of Vascular

Inflammation in Rheumatoid Arthritis. Life Sciences,

93(8), pp. 307–312.

Roberts, J. D., Roberts, M. G., Tarpey, M. D., Weekes, J.

C., & Thomas, C. H., 2015. The effect of a

decaffeinated green tea extract formula on fat

oxidation, body composition and exercise performance.

Journal of the International Society of Sports Nutrition,

12.

Rohman, M. S., 2011. Patogenesis dan Terapi Sindroma

Metabolik. Jurnal Kardiologi Indonesia, 28(2), pp. 86–

94.

Saifur Rohman, M., Lukitasari, M., Adi Nugroho, D.,

Nashi, W., Ida Panca Nugraheini, N., & Wahyu

Sardjono, Teguh., 2017. Development of an

Experimental Model of Metabolic Syndrome in

Sprague Dawley Rat. Research Journal of Life Science,

4(1), pp. 76–86.

Wu, L.-Y., Juan, C.-C., Hwang, L. S., Hsu, Y.-P., Ho, P.-

H., & Ho, L.-T., 2004. Green tea supplementation

ameliorates insulin resistance and increases glucose

transporter IV content in a fructose-fed rat model.

European Journal of Nutrition, 43(2), pp. 116–124.

Zhang, Z. F., Li, Q., Liang, J., Dai, X. Q., Ding, Y., Wang,

J. B., & Li, Y., 2010. Epigallocatechin-3-O-gallate

(EGCG) protects the insulin sensitivity in rat L6 muscle

cells exposed to dexamethasone condition.

Phytomedicine: International Journal of Phytotherapy

and Phytopharmacology, 17(1), pp. 14–18.

JIMC 2020 - 1’s t Jenderal Soedirman International Medical Conference (JIMC) in conjunction with the Annual Scientiﬁc Meeting

(Temilnas) Consortium of Biomedical Science Indonesia (KIBI )