【摘要】 目的研究AMPK活化对INS-1细胞胰岛素释放的作用及其可能机制。方法体外培养INS-1细胞株,观察不同浓度(0.2,0.5和1.0mmol·L-1)AICAR(AMPK激动剂)作用不同时相点(8,12和24h)对细胞内胰岛素含量及高糖刺激的胰岛素释放的影响;AICAR(0.5mmol·L-1)与Compound C(10μmol·L-1,AMPK阻断剂)单独或共同作用INS-1细胞8h,采用两种PCR(RT-PCR和实时定量PCR)方法检测PPARα基因转录水平的变化,免疫沉淀检测PPARα蛋白表达。结果与正常对照组比较,AICAR(0.2,0.5和1mmol·L-1)作用8,12和24h,均抑制INS-1细胞高糖刺激的胰岛素释放及细胞内胰岛素含量。同时,AICAR诱发的AMPK活化能增强PPARα mRNA和蛋白水平的表达。结论AICAR诱导的AMPK活化可能通过调节PPARα表达抑制INS-1细胞高糖刺激的胰岛素释放。更多还原

【Abstract】 Aim To explore the effect of AMPK activation on insulin release in INS-1 cells and its possible mechanisms.Methods INS-1 cell line was cultured in vitro.The cells were treated with different concentrations(0.2,0.5 and 1.0 mmol·L-1)of AICAR(AMPK activator)for 8,12,or 24 h.High glucose-stimulated insulin release and intracellular insulin content were assayed by RIA,respectively.INS-1 cells were incubated in the presence and Absence of AICAR(0.5 mmol·L-1),treated with and without Compound C(10 μmol·L-1,AMPK inhibitor)for 8 h,the level of PPARα gene transcription was measured by using both RT-PCR and real-time PCR,and the expression of PPARα protein was analyzed by immunoprecipitation.Results Compared with control group,0.2,0.5,or 1.0 mmol·L-1 AICAR decreased the high glucose-stimulated insulin release and intracellular insulin content when the cells were treated for 8,12,or 24 h.Meanwhile,AMPK activation up-regulated the levels of PPARα mRNA and protein expression.Conclusion AMPK activation induced by AICAR can inhibit the high glucose-stimulated insulin release in INS-1 cells through regulating the expression of PPARα.更多还原