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探讨胡椒碱对胰岛素抵抗(insulin resistance,IR)细胞模型糖代谢紊乱的作用及干预AMPK信号通路上游靶点的分子机制。通过脂肪乳诱导HepG2细胞构建胰岛素抵抗模型。葡萄糖氧化酶(GOD-POD)法检测各组葡萄糖消耗量;酶联免疫吸附测定(ELISA)法检测上清液中脂联素(adiponectin,APN)、瘦素(leptin,LEP)的水平;荧光定量PCR法检测APN,LEP mRNA的表达水平,Western blotting法检测AMPKα,р-AMPKα,瘦素受体(LepR),脂联素受体1(AdipoR1)和脂联素受体2(AdipoR2)蛋白的表达。结果显示,胡椒碱与降糖药罗格列酮、AMPK激动剂AICAR均能明显提高胰岛素抵抗细胞模型的葡萄糖消耗量,升高脂联素水平、增加脂联素mRNA、脂联素受体1蛋白的表达,也增加AMPKαmRNA和р-AMPKα蛋白表达;同时降低瘦素mRNA和瘦素受体蛋白的表达。结果表明,胡椒碱能显著改善胰岛素抵抗细胞模型糖代谢紊乱,其作用机制可能通过调控上游脂联素和瘦素的表达,从而激活AMPK信号通路。
To investigate the effect of piperine on the disorder of glucose metabolism in insulin resistance (IR) cell model and the molecular mechanism of intervention on the upstream target of AMPK signaling pathway. HepG2 cells were induced to model insulin resistance by lipid emulsion. Glucose oxidase (GOD-POD) method was used to detect the glucose consumption in each group. The levels of adiponectin (APN) and leptin (LEP) in the supernatant were detected by enzyme linked immunosorbent assay (ELISA) The expression of APN and LEP mRNA was detected by quantitative PCR. The protein levels of AMPKα, р-AMPKα, LepR, AdipoR1 and AdipoR2 were detected by Western blotting. expression. The results showed that piperine and hypoglycemic agents rosiglitazone, AMPK agonist AICAR can significantly improve the insulin resistance cell model of glucose consumption, increased adiponectin levels, increased adiponectin mRNA, adiponectin receptor 1 Protein expression, but also increased AMPKα mRNA and р-AMPKα protein expression; at the same time reduce leptin mRNA and leptin receptor protein expression. The results showed that piperine could significantly improve the disorder of glucose metabolism in insulin resistance cell model. Its mechanism may activate the AMPK signaling pathway by regulating the expression of upstream adiponectin and leptin.