Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3<italic style="font-style: italic">β</italic> Signal Transduction Pathway

Zi-yang DAI; Yu-shan DONG; Pei-jie DING; Yu-yang YANG; Chen-guang DU; Ya WANG; Cai-jian HE

doi:10.13422/j.cnki.syfjx.20191204

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Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway

Pharmacology | 更新时间：2021-02-09

- Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 12, Pages: 77-82(2019)
- 作者机构：
  
  华北理工大学，河北　唐山　 063210
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20191204
  CLC： R2-0; R22; R285.5
- Received：23 November 2018，
  
  Published Online：04 March 2019，
  
  Published：20 June 2019
- 稿件说明：
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Zi-yang DAI, Yu-shan DONG, Pei-jie DING, et al. Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 77-82.
DOI：

Zi-yang DAI, Yu-shan DONG, Pei-jie DING, et al. Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(12): 77-82. DOI： 10.13422/j.cnki.syfjx.20191204.

摘要

目的：

观察葛根素对胰岛素抵抗HepG2细胞内磷脂酰肌醇3激酶(PI3K)，蛋白激酶B(Akt)，糖原合成酶激酶-3

(GSK-3

)信号分子的影响。

方法：

通过0.5 mmol·L

－1

棕榈酸联合9×10

－4

U·L

－1

胰岛素孵育24 h诱导HepG2胰岛素抵抗细胞模型，噻唑蓝(MTT)比色法测定细胞存活率以确定葛根素给药浓度。实验设正常组、模型组和葛根素各剂量组(40，80，160，320 μg·L

－1

)。葡萄糖检测试剂盒检测细胞培养液上清葡萄糖的含量，酶联免疫吸附测定(ELISA)检测细胞培养液上清中肿瘤坏死因子-

(TNF-

)，白细胞介素-6(IL-6)的含量，肝糖原测定试剂盒测定HepG2细胞内肝糖原含量，蛋白免疫印迹法(Western blot)检测细胞内PI3K，Akt，磷酸化(p)-Akt，GSK-3

，p-GSK-3

的蛋白表达水平。

结果：

与正常组比较，模型组HepG2细胞葡萄糖消耗率明显下调(

0.05)，细胞内糖原含量减少(

0.05)，细胞培养液上清中TNF-

和IL-6的含量增加(

0.05)，PI3K和p-Akt蛋白表达下降(

0.05)，GSK-3

蛋白表达增加(

0.05)；与模型组比较，葛根素可呈剂量依赖性增加细胞葡萄糖消耗率(

0.05)，增加细胞内肝糖原含量(

0.05)，降低细胞培养液上清中TNF-

和IL-6的含量(

0.05)，增加PI3K蛋白表达，上调Akt蛋白磷酸活化水平(

0.05)，下调GSK-3

蛋白表达并增强其磷酸失活水平(

0.05)。

结论：

葛根素可通过增强PI3K/Akt/GSK-3

信号转导过程，增加肝细胞内糖原含量从而改善HepG2细胞的胰岛素抵抗状态。

Abstract

Objective:

To observe the effect of puerarin on phosphatidylinositol 3-kinase (PI3K)

protein kinase B (Akt) and glycogen synthase kinase-3

(GSK-3

) in insulin resistant HepG2 cells.

Method:

HepG2 cells were treated with palmitic acid 0.5 mmol·L

-1

and insulin 9×10

-4

U·L

-1

to induce insulin resistant condition for 24 h. Cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay to determine the concentration of puerarin. This experiment included normal control group

model control group and puerarin groups of different doses (40

160

320 μg·L

-1

). Glucose detection kit was used to detect the content of glucose in cell culture supernatant. Tumor necrosis factor alpha (TNF-

) and interleukin-6 (IL-6) levels in supernatant of cell culture medium were detected by enzyme-linked immunosorbent assay (ELISA). Hepatic glycogen assay kit was used for detecting the hepatic glycogen content in HepG2 cells. Western blot was applied to detect protein expression levels of PI3K

Akt

p-Akt

GSK-3

and p-GSK-3

Result:

Compared with those in the normal control group

the glucose consumption rate was significantly down-regulated in HepG2 cells in the model control group (

0.05)

the intracellular glycogen content was decreased (

0.05)

the content of TNF-

and IL-6 were increased in supernatant of cell culture medium (

0.05)

PI3K protein level was decreased

Akt phosphorylation level was down-regulated (

0.05)

and GSK-3

protein expression was up-regulated (

0.05). Compared with those in the model control group

in the puerarin groups

the glucose consumption rate was increased in a dose-dependent manner (

0.05)

the intracellular glycogen content was increased

the contents of TNF-

and IL-6 were reduced in supernatant of cell culture medium (

0.05)

PI3K protein expression was enhanced

Akt phosphorylation level was up-regulated (

0.05)

GSK-3

protein expression was down-regulated

but its phosphorylation inactivation was increased (

0.05).

Conclusion:

Puerarin alleviates the insulin resistance of HepG2 cells by strengthening the PI3K/Akt/GSK-3

signal transduction process and increasing the glycogen content in hepatocytes.

关键词

Keywords

references

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Effect of Puerarin in Reducing Insulin Resistance in HepG2 Cells via PI3K/Akt/GSK-3β Signal Transduction Pathway

DOI：10.13422/j.cnki.syfjx.20191204

摘要

Abstract

关键词

Keywords

references