复方贞术调脂方对HepG2细胞胰岛素抵抗的作用及机制研究

Research of FTZ on the function and mechanism of insulin resistance in HepG2 cells

目的观察复方贞术调脂方(FTZ)对胰岛素抵抗(IR)HepG2细胞的作用,为开拓FTZ的治疗范围,阐明FTZ调节糖脂代谢的机制提供实验依据。方法用高浓度胰岛素诱导HepG2细胞使其产生胰岛素抵抗,用高、中、低3个剂量的FTZ干预后,葡萄糖氧化酶法检测HepG2细胞培养液上清液中葡萄糖的含量,实时荧光定量PCR检测HepG2细胞胰岛素信号PI-3Kp85 mRNA的表达,Western blot检测HepG2细胞胰岛素信号转导蛋白IRS1表达。结果模型细胞培养基上清液中葡萄糖含量高于正常细胞(P<0.05)。给予FTZ(1、25、100μg/mL)后,HepG2细胞培养基上清液中葡萄糖含量均低于模型细胞葡萄糖含量(P<0.05)。胰岛素抵抗细胞与正常细胞相比PI-3Kp85 mRNA和IRS1的蛋白表达显著降低(P<0.05)。给予FTZ干预后,与胰岛素抵抗细胞相比PI-3Kp85 mRNA和IRS1的蛋白表达显著增加(P<0.05)。结论FTZ可改善胰岛素抵抗HepG2细胞对葡萄糖的摄取。其改善胰岛素抵抗的作用机制之一可能是通过上调胰岛素信号PI-3Kp85 mRNA和IRS1蛋白质在胰岛素抵抗HepG2细胞的表达。

Objective To investigate the effect and mechanism of Fufang Zhenzhu Tiaozhi formula （FTZ） on insulin-resistant HepG2 cells for further development and clinical application.Methods HepG2 cells were induced with high insulin as a model of insulin resistance and treated with FTZ at three different dosages. The levels of glucose content, PI3K p85 mRNA and IRS1 protein were measured by GOD （glucose oxidase） method, real-time PCR and west blotting, respeetively. Results Tile glucose content ill cultured insulin-resistant HepG2 cells was obviously increased, which was decreased when treated with FTZ. Compared with the eontrol group,the expression of IRS1 protein and PI-3K p85 mRNA was decreased in insulin-resistant HepG2 cells （P〈0.05）.Treatment with FTZ increased IRS1 protein and PI-3K p85 mRNA expression in insulin-resistant HepG2 cells （P〈0.05）. Conclusion FTZ can improve glucose uptake in insulin-resistant HepG2 cells,whieh may be related with up-regulation of the expression of PI-3K p85 and IRS1 in the insulin IRS-PI3K-Akt pathway.