AMPK调控Ca2＋内流对高糖诱导内皮细胞凋亡的作用及其机制研究  

Effects of AMPK on high glucose stimulated apoptosis of endothelial cells via regulation of calcium influx

作　　者：卢婷[1] 郜攀[1] 司良毅[1] 赵坤[1] 

机构地区：[1]重庆第三军医大学西南医院老年病科,400038

出　　处：《解放军医学杂志》2015年第10期773-777,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81370446、81370007)~~

摘　　要：目的观察AMP依赖的蛋白激酶(AMPK)对高糖刺激内皮细胞凋亡的抑制作用,并初步探讨其机制。方法体外培养MS-1内皮细胞株,分别用AMPK激动剂、AMPK抑制剂、钙库依赖性钙离子通道(SOCC)抑制剂2-APB和(或)高糖处理,另设对照组(未经任何方式干预)。采用TUNEL法检测细胞凋亡情况,激光共聚焦显微镜检测细胞内钙离子(Ca2+)内流,Western blotting检测SOCC蛋白Stim1和Orai1的表达。结果与对照组比较,高糖能够明显诱导内皮细胞凋亡,增加Stim1和Orai1蛋白表达(P<0.05)。与高糖组比较,AMPK抑制剂+高糖能够明显增强高糖诱导的内皮细胞的凋亡(P<0.05),而AMPK激动剂+高糖能够明显抑制高糖诱导的内皮细胞凋亡,并降低Stim1和Orai1蛋白表达(P<0.05)。与对照组比较,高糖能够明显诱导内皮细胞Ca2+内流;与高糖组比较,2-APB+高糖能够明显抑制高糖诱导的内皮细胞Ca2+内流,并阻断高糖对内皮细胞凋亡的诱导作用,而AMPK激动剂能够明显抑制高糖诱导的内皮细胞Ca2+内流。结论 AMPK能够通过降低Stim1和Orai1蛋白的表达,抑制SOCC介导的Ca2+内流,进而阻断高糖刺激的内皮细胞凋亡,对内皮细胞功能起重要的保护作用。Objective To investigate the inhibitory effect of adenosine monophosphate （AMP）-dependent protein kinase （AMPK） on high glucose-stimulated endothelial cell apoptosis and its mechanism. Methods MS-1 endothelial cells were cultured in vitro, and they were treated with AMPK agonist, AMPK inhibitor, 2-APB （a blocker of store operated Ca2＋ channel （SOCC）） and （or） high glucose, and a control group without any intervention were set up. TUNEL assay was performed to determine apoptotic cells. Laser scanning confocal microscopy was used to assess the Ca2＋ influx into cells, and Western-blotting was performed to determine the expressions of Stiml and Orail of the store operated Ca2＋ channel （SOCC） proteins. Results Apoptosis of endothelial cells was induced significantly, and the expressions of Stiml and Orail were upregulated in high glucose group compared with that in control group （P〈0.05）. The rate of apoptosis of high glucose-induced endothelial cell was found to be increased in AMPK inhibitor group and decreased in AMPK agonist group, and the expressions of Stiml and Oral1 were found to be down- regulated in AMPK agonist group as compared with that in high glucose group （P〈0.05）. Compared with the control group, high glucose stimulation significantly induced the Ca2＋ influx to endothelial cells; compared with high glucose group, 2-APB significantly inhibited high glucose-induced Ca2＋ influx to endothelial cells, and blocked the inducing effect of high-glucose on endothelial cell apoptosis. Compared with high glucose group, AMPK agonist significantly inhibited high glucose-induced cell Ca2＋ influx. Conclusion By reducing the expressions of Stiml and Orail, AMPK may inhibit SOCC-mediated Ca2＋ influx, and block the high glucose-stimulated endothelial cell apoptosis, thus play an important protective role in sustaining endothelial cell function.

关 键 词：AMP活化蛋白激酶类 钙离子载体 高血糖症 钙库依赖性钙离子通道 内皮细胞 

分 类 号：R345[医药卫生—基础医学]