异鼠李素调控AKT-FOXO1通路改善胰岛素抵抗HepG2细胞糖代谢作用机制

The Mechanism of Isorhamnetin Regulating AKT-FOXO1 Pathways on Improve the Insulin Resistance Model of HepG2 Cells

目的:研究异鼠李素通过AKT-FOXO1信号通路改善Hepg2细胞胰岛素抵抗机制。方法:使用四甲基偶氮唑盐微量酶反应比色(MTT)法筛选异鼠李素安全剂量;通过高浓度胰岛素处理诱导Hepg2细胞胰岛素抵抗(IR)模型;使用葡萄糖氧化酶法测得培养基中葡萄糖含量,衡量异鼠李素对胰岛素抵抗Hepg2细胞葡萄糖代谢水平影响;通过Western blot检测细胞中AKT(蛋白激酶B),p-AKT(磷酸化的蛋白激酶B),FOXO1(叉头转录因子O亚家族1),p-FOXO1(磷酸化的叉头转录因子亚家族1),G6pase(六磷酸葡萄糖酶),PEPCK(磷酸烯醇式丙酮酸激酶)这六个蛋白表达水平的变化。结果:异鼠李素对Hepg2细胞的安全剂量为1~3.8μg/mL;与模型组相比,低浓度异鼠李素显著的增加了胰岛素抵抗Hepg2葡萄糖消耗量(P<0.05),显著促进了胰岛素抵抗Hepg2细胞的AKT、FOXO1蛋白磷酸化(P<0.05),从而显著抑制了G6pase和PEPCK的蛋白表达(P<0.05),其作用效果与二甲双胍相当。结论:异鼠李素可有效改善胰岛素抵抗Hepg2细胞,并通过调节AKT-FOXO1信号通路达到这一作用效果。

Objective:To research the mechanism of isorhamnetin in treating insulin resistance,insulin-resistant Hepg2 cells were treated with isorhamnetin.Method:The MTT method was used to screen the safe dose of isorhamnetin.Hepg2 cells insulin resistance(IR)model was induced by high-concentration insulin treatment.The glucose oxidase method was used to measure the glucose content of the medium,to measure the effect of isorhamnetin on glucose metabolism in insulin resistant Hepg2 cells.The expression levels of AKT,p-akt,FOXO1,p-foxo1,G6pase,PEPCK and six proteins were detected by Western blot.Results:The safe dose of isorhamtin to Hepg2 cells was 1-3.8μg/mL.Isorhamnetin effectively increased insulin resistance Hepg2 glucose consumption(P<0.05).Phosphorylation of AKT and FOXO1 proteins expression in insulin resistant(P<0.05)Hepg2 cells was promoted and the protein expression of G6pase and PEPCK was inhibited(P<0.05).The effect of isorhamnetin was comparable to metformin.Conclusion:Isorhamnetin can effectively improve insulin resistance of Hepg2 cells and achieve this effect by regulating the AKT-FOXO1 signaling pathway.