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。
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 acti- vator）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 i.Lmol·L^-1, AMPK inhibitor）for 8 h,the level of PPARα gene transcription was measured by using both RT-PCR and realtime PCR,and the expression of PPARα protein was an-alyzed 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 upregulated 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.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2008年第2期183-187,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30670994)
山东省自然科学基金资助项目(NoY2005C03)
山东省卫生厅资助项目(No2005HZ061)
