自噬对高糖诱导的人冠状动脉内皮细胞凋亡的保护作用  

Protective Effect of Autophagy on Human Coronary Artery Endothelial Cell Apoptosis Induced by High Glucose

作　　者：刘宇[1] 曹清心[2] 石屹城[1] 崔玲[1] 陈启[1] 于曼丽[1] 赵仙先[1] 

机构地区：[1]第二军医大学附属长海医院心内科,上海200433 [2]第二军医大学附属长海医院中医科,上海200433

出　　处：《现代生物医学进展》2015年第12期2201-2204,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81370266;81170223);上海市自然科学基金项目(134119a0301;13ZR1450300)

摘　　要：目的:研究自噬对高糖诱导的人冠状动脉内皮细胞凋亡的影响。方法:将人冠状动脉内皮细胞,分别用常规培养基(正常对照组)、含30 mmol/L D-葡萄糖的高糖培养基(高糖组)、高糖培养基合并雷帕霉素(Rapamycin,RAPA;100 nmol/L)干预(RAPA组)和高糖培养基合并3-甲基腺嘌呤(3-Methyladenine,3-MA,5 mmol/L)干预(3-MA组)培养。利用CCK-8法检测细胞生长活力,使用流式细胞术检测细胞凋亡水平,western blot检测细胞自噬标记蛋白(Beclin1)的表达水平。结果:(1)高糖溶液刺激内皮细胞24 h后,细胞生长活力为正常组的55.0%(P<0.01),自噬标记蛋白Beclin1的表达水平明显增加,凋亡水平为正常组的2.0倍;(2)与高糖组相比,RAPA组细胞生长活力明显增加,Beclin1的表达明显升高(P<0.01),凋亡水平为高糖组的70.1%;(3)与高糖组相比,3-MA组细胞生长活力明显减少,Beclin1的表达明显降低(P<0.01),凋亡水平为高糖组的1.42倍。结论:细胞自噬可能对高糖诱导的人冠状动脉内皮细胞具有凋亡保护作用。Objective： To study the protective effect of autophagy on endothelial cells apoptosis induced by high glucose. Methods： Human coronary artery endothelial cells cultured by conventional media （Control group）, conventional media contraining 30 mmol/L D-glucose （High-glucose group）, high-glucose media plus 100 nmol/L Rapamycin （RAPA group）, or high-glucose media plus 5mmol/L 3-Methyladenine （3-MA group）. Cells vitality and apoptosis were measured by CCK-8 assay and flow cytometry. The autophagy marking protein （Beclinl） expression was determined by western blot. Results： （1） Endothelial cell viability decreased significantly after high-glucose （30 mmol/L） stimulation, with 55.0% to the control group （P〈0.01）. Beclinl expressed increased, and the level of cell apoptosis was significanly increased, with 2 times to the control group. （2） Compared with high-glucose group, the cell viability and the beclinl expression of RAPA group were both increased （P〈0.01）. The level of cell apoptosis was significanly decreased, with 70.1% to the high-glucose group. （3） Compared with high-glucose group, the cell viability and the beclinl expression of RAPA group were both decreased （P〈0.01）. The level of cell apoptosis was significanly increased, with 1.42 times to the high-glucose group. Conclusion： Cell autophagy might have an apoptosis-protective effect on the human coronary artery endothelial cells induced by high-glucose.

关 键 词：自噬 内皮细胞 高糖 凋亡 保护 

分 类 号：Q75[生物学—分子生物学] R331.3[医药卫生—人体生理学]