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目的观察二甲双胍对IR状态的小鼠胚胎成纤维(3T3-L1)细胞中腺苷酸活化蛋白激酶(AMPK)及葡萄糖转运子4(GluT4)mRNA表达的影响。方法诱导分化3T3-L1前脂肪细胞为成熟脂肪细胞并用油红O染色证明。用地塞米松诱导建立3T3-L1细胞IR模型,以葡萄糖氧化酶法测定不同时段细胞培养基中剩余葡萄糖浓度,以确定IR模型的建立。给予二甲双胍进行药物干预后,测定细胞培养基中剩余葡萄糖浓度并运用实时荧光定量PCR技术检测细胞AMPK和GluT4的mRNA基因水平。结果药物干预组给予不同浓度二甲双胍后,细胞培养基中剩余葡萄糖浓度均较模型组降低(P<0.01)。药物干预组AMPK、GluT4mRNA水平较模型组均增加(P<0.01)。结论二甲双胍上调处于IR状态的3T3-L1细胞中AMPK和GluT4mRNA水平,增加细胞对葡萄糖利用。
Objective To observe the effects of metformin on the expression of AMPK and GluT4 mRNA in the mouse embryonic fibroblast (3T3-L1) cells in the IR state. Methods Induction of differentiation of 3T3-L1 preadipocytes into mature adipocytes was confirmed by oil red O staining. The IR model of 3T3-L1 cells was induced by dexamethasone, and the remaining glucose concentration in the cell culture medium was determined by glucose oxidase method to determine the establishment of IR model. Metformin was given drug intervention, the remaining glucose concentration in the cell culture medium was determined and mRNA levels of AMPK and GluT4 in cells were measured by real-time fluorescence quantitative PCR. Results After treatment with different concentrations of metformin, the concentration of glucose in the cell culture medium was lower than that of the model group (P <0.01). Compared with the model group, the levels of AMPK and GluT4 mRNA in the intervention group increased (P <0.01). Conclusion Metformin up-regulates AMPK and GluT4 mRNA levels in 3T3-L1 cells that are in IR state, increasing cell glucose utilization.