解偶联蛋白2对高糖高脂高尿酸诱导的心肌细胞凋亡的作用及机制  

Effect and mechanism of uncoupling protein 2 on cardiomyocyte apoptosis induced by high glucose,high lipid and high uric acid

作　　者：田玥 杨怡[1,2] 贺磊[2] 侯娟妮 杜劲[2] 陈莎[2] 王挺[2] 裴海峰[2] 杨永健[1,2] 

机构地区：[1]西南医科大学临床医学院,四川泸州646000 [2]成都军区总医院心血管内科,成都610083

出　　处：《解放军医学杂志》2017年第6期520-525,共6页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81500208);四川省科技厅杰出青年基金(2017JQ0012);四川省科技支持计划项目(2015JY0277);博士后医院管理基金(41732BA)~~

摘　　要：目的研究解偶联蛋白2(UCP2)在高糖高脂高尿酸小鼠心肌细胞(MCM)中的作用及机制。方法以25mmol/L高糖培养基+300μmol/L软脂酸钠干预MCM 18h模拟合并高脂血症的2型糖尿病组(高糖+高脂组),在糖尿病组的基础上再用1500μmol/L尿酸干预18h模拟2型糖尿病合并高尿酸组(高糖+高脂+高尿酸组)。随后,采用UCP2抑制剂Genipin抑制心肌线粒体UCP2表达,进一步将2型糖尿病合并高尿酸组分为溶媒对照组、Genipin组。研究UCP2在高糖高脂合并高尿酸心肌细胞损伤的机制时,再分为Genipin组、Genipin+N-乙酰半胱氨酸(NAC)组。采用流式细胞仪检测各组心肌细胞的凋亡水平,q-PCR法和Western blotting检测心肌细胞中UCP2的表达情况,DHE染色及ELISA法检测活性氧簇(ROS)水平。结果与高糖+高脂组比较,高糖+高脂+高尿酸组心肌细胞凋亡明显增加(P<0.05),心肌细胞中UCP2的表达水平明显降低,同时ROS的水平明显上升(P<0.05)。采用Genipin抑制UCP2的表达水平后,高糖高脂高尿酸干预后心肌细胞凋亡水平及ROS水平升高更加明显(P<0.05)。在此基础上应用抗氧化剂NAC后,高尿酸所诱导的心肌细胞凋亡及ROS升高被明显逆转(P<0.05)。结论 UCP2可以通过抑制氧化应激缓解高糖高脂合并高尿酸所诱导的心肌细胞凋亡。Objective To investigate the effects of uncoupling protein 2 （UCP2） on the myocardial cells of mice with type 2 diabetes mellitus combined with hyperuricemia （HUA）, and clarify the mechanism thereof. Methods The mouse cardiac myocytes （MCM） cultured with 25mmol/L high glucose （HG） medium were divided into two groups： HG plus 300μmol/L sodium palmitate for 18 hours as high glucose and high fat （HG＋HF） group, and HG＋HF plus 1500μmol/L uric acid （UA） for 18 hours as HG＋HF＋HUA group. Then the myocardial cells in HG＋HF＋HUA group, by use or not use UCP2 inhibitor genipin, were further divided into two groups： vehicle group and genipin group. In order to verify the mechanism of UCP2 in myocardial cells injury caused by high glucose, high lipid and high uric acid, the myocardial cells were divided again into genipin group and genipin＋N-acetylcysteine （NAC） group. Accordingly, the apoptosis of myocardial cells were measured by flow cytometry at specific time, the mRNA and protein expressions of UCP2 were determined by q-PCR and Western blotting, and the levels of reactive oxygen species （ROS） were detected by DHE staining and ELISA. Results The apoptosis rate of myocardial cells increased obviously, and the expression levels of UCP2 decreased and of ROS elevated significantly in HG＋HF＋HUA group than in HG＋HF group （P〈0.05）. As the expression levels of UCP2 decreased by genipin intervention, the apoptosis rate of myocardial cells and ROS level in HG＋HF＋HUA group increased more obviously （P〈0.05）. In contrast, such an effect was reversed by the application of antioxidants NAC （P〈0.05）. Conclusion UCP2 can inhibit oxidative stress and alleviate the apoptosis of myocardial cells induced by high glucose, high fat and high uric acid.

关 键 词：高尿酸血症 糖尿病 解偶联蛋白2 氧化应激 心肌细胞 

分 类 号：R543.5[医药卫生—心血管疾病]