[关键词]
[摘要]
目的 通过体外实验探讨羁糖脂改善2型糖尿病胰岛素抵抗(IR)的作用,并初步探明其作用机制。方法 采用1×10-7 mol/L胰岛素诱导HepG2细胞,建立IR细胞模型;MTT法检测羁糖脂对HepG2细胞的增殖抑制率;葡萄糖氧化酶-过氧化物酶(GOD-POD)法检测羁糖脂对HepG2 IR细胞葡萄糖吸收的影响;Western blotting法检测胰岛素受体底物1磷酸化丝氨酸p-IRS-1(Ser307)、磷酸酰肌醇-3-激酶(PI3K)、葡萄糖转运体-4(GLUT-4)的表达。结果 HepG2细胞置于含10-7胰岛素培养液孵育24 h,与对照组比较,葡萄糖吸收水平下降,表明建模成功;30~120 μg/mL羁糖脂均能显著增加IR细胞葡萄糖吸收率,显著上调GLUT-4、PI3K蛋白表达水平,显著下调IRS-1 Ser307磷酸化水平。结论 羁糖脂能有效增强HepG2 IR细胞对葡萄糖的消耗能力,推测与上调GLUT-4、PI3K蛋白表达,抑制IRS-1丝氨酸磷酸化相关。
[Key word]
[Abstract]
Objective To evaluate the effect of Ji Tang Zhi on glucose metabolism in insulin resistance (IR) HepG 2 cell line, and to explore the related mechanism. Methods The HepG2 cells were incubated in culture medium addition of 10-7 mol/L insulin for 24 h to establish the IR cell model. Effect of Ji Tang Zhi on rate of glucose absorption in HepG2 cell was detected by the method of glucose oxidase-peroxidase (GOD-POD). We performed an MTT assay to determine cytotoxicity effects of Ji Tang Zhi on HepG2 cell line. The expression of p-IRS-1 Ser307, PI3K and GLUT-4 were detected by Western blotting. Results Incubated with 10-7 mol/L insulin for 24 h, the insulin resistance cell model had been built. Compared with model group, the rate of glucose absorption of cell treated with JTZ (30 ~120 μg/mL) was significantly improved. According to model cells, the expression of GLUT-4 and PI3K decreased significantly compared to control cells. While the expression of p-IRS-1 Ser 307 was inhibited and GLUT-4 and PI3K expression were increased in IR cells after treated with JTZ (30 ~120 μg/mL). Conclusion JTZ exert beneficial effects on hyperglycosemia in IR cell line possibly through regulating the levels of GLUT-4, p-IRS-1 Ser307 and PI3K in HepG2 cell.
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[基金项目]
淮安市科技局基金资助项目(HAN2015025)