葛根芩连汤含药血清对HepG2肝细胞胰岛素抵抗模型糖代谢的调节作用

Effects of Gegen Qinlian Tang-containing Serum on Glycometabolism in Insulin-resistance HepG2 Cell Line

摘要:

目的: 探讨葛根芩连汤含药血清对HepG2肝细胞胰岛素抵抗模型糖代谢的调节作用。方法: 大鼠分别按含生药 6 g·kg^-1剂量ig葛根芩连汤,空白组ig给予等量蒸馏水,按3 mg·kg^-1剂量ig吡格列酮,2次/d,连续7 d,末次给药1 h后,心 脏穿刺取血制备葛根芩连汤、空白和吡格列酮含药血清。采用MTT法检测葛根芩连汤含药血清对细胞增殖的影响;HepG2细胞以含10%胎牛血清(FBS)的DMEM培养基传代培养,将处对数生长期细胞消化后,用含2%FBS DMEM培养基调整细胞密度为10⁶个/mL, 以10 mg·L^-1胰岛素处理24 h,复制HepG2细胞胰岛素抵抗模型,葡萄糖氧化酶法检测葛根芩连汤含药血清对HepG2细胞葡萄糖消耗的影响;蒽酮法检测细胞内糖原含量;乳酸脱氢酶偶联比色法测定肝细胞磷酸烯醇式丙酮酸羧激酶(PEPCK)的活性。结果: 与正常组比较,模型组葡萄糖消耗量明显下降(P<0.01),糖原含量显著降低(P<0.05), 而PEPCK活性升高 (P<0.05);8%葛根芩连汤含药血清作用HepG2细胞胰岛素抵抗模型24 h能显著增加葡萄糖消耗量(P<0.05),且葛根芩连汤含药血清(8%,16%)能够增加糖原含量,葛根芩连汤含药血清(4%,8%,16%)能够降低PEPCK活性(P<0.05)。结论: 葛根芩连汤含药血清可调节肝糖代谢,进而可改善肝细胞胰岛素抵抗。

Abstract:

Objective: To investigate the effects of Gegen Qinlian Tang-containing serum (GGQLT-CS) on glycometabolism in insulin-resistance HepG2 cell line. Method: GGQLT-CS was prepared.MTT was used to detect the effects of GGQLT-CS on cell proliferation;the model of insulin resistance was established,the effect of GGQLT-CS on the glucose consumption of HepG2 cells was observed in insulin resistance through the method of glucose oxidase, detecting phosphoenolpyruvate carboxykinase (PEPCK) activity was detected with the method of the lactate dehydrogenase coupled assay, and the anthrone method was used to detect intracellular glycogen content. Result: GGQLT-CS could promote glucose consumption of HepG2 cell in insulin resistance, reduce PEPCK activity, and increase hepatic glycogen content. Conclusion: GGQLT-CS can regulate glucose metabolism of HepG2 cells, and improvehepatic insulin resistance.