高尿酸通过激活钙蛋白酶加重缺血/缺氧条件下小鼠心肌细胞凋亡  

Hyperuricemia aggravates cardiomyocytes apoptosis induced by ischemia/hypoxia via activating calpains pathway

作　　者：贺磊 杨怡[2] 田玥[2] 邓晶晶[3] 李秀川[2] 裴海峰[2] 杨大春[1,2] 

机构地区：[1]第三军医大学研究生管理大队,重庆400038 [2]成都军区总医院心内科 [3]成都医学院药学院

出　　处：《临床心血管病杂志》2017年第10期1004-1008,共5页Journal of Clinical Cardiology

基　　金：国家自然科学基金(No:81500208);四川省科技厅杰出青年基金(No:2017JQ0012);四川省医学科研青年创新课题(No:Q14005)

摘　　要：目的:研究高尿酸对缺血/缺氧条件下心肌细胞的影响及钙蛋白酶(Calpains)在其中的作用。方法:将小鼠心肌细胞系分为3组:0μmol/L尿酸+缺血/缺氧组(单纯缺血/缺氧组);600μmol/L尿酸+缺血/缺氧组;1 200μmol/L尿酸+缺血/缺氧组,通过流式细胞技术检测各组心肌细胞凋亡比例,并进一步将凋亡比例明显增加的实验组分为:1 200μmol/L尿酸+缺血缺氧组;MDL28170+1 200μmol/L尿酸+缺血缺氧组。采用DHE染色法与ELISA试剂盒测定心肌细胞中活性氧簇(ROS)水平,q-PCR法及Western blot检测心肌细胞中Calpain1、Calpain2、Calpastatin的表达情况。结果:高尿酸可明显增加缺血/缺氧状态下心肌细胞凋亡水平(P<0.05)。与单纯缺血/缺氧组相比,高尿酸显著增加了Calpain1/2的表达,明显抑制了Calpastatin的表达(P<0.05),同时显著增加了心肌细胞中ROS的水平(P<0.05),而在加入MDL28170干预后,心肌细胞的ROS水平及凋亡比例均有所降低(P<0.05)。结论:高尿酸通过激活Calpain所介导的氧化应激反应加重缺血/缺氧状态下心肌细胞凋亡。Objective：To investigate the effects of elevated uric acid on myocardial cells in ischemic condition and to clarify the role of calpains in it.Method：The myocardial cell line（MCM）was divided into three groups：0μmol/L uric acid group,600μmol/L group and 1 200μmol/L uric acid,all of groups treated with ischemia/hypoxia.Furthermore,the intervention group of much higher apoptosis rate was divided into two groups：1 200μmol/L uric acid with ischemia/hypoxia group and 1 200μmol/L uric acid with ischemia/hypoxia group incubated with Calpain inhibitor MDL28170（6mmol/L）for 24 hours.Accordingly,the apoptosis of cardiomyocytes were measured by flow cytometer,the levels of reactive oxygen species（ROS）were detected by DHE staining and ELISA kits,the expressions of Calpain1/Calpain2/Calpastatin were determined by q-PCR and Western blot.Result：The apoptotic ratio of myocardial cells gradually increased with elevated concentration of uric acid（P〈0.05）.Moreover,compared with control group,hyperuricemia obviously stimulated the expression of calpains in cardiomyocytes（P〈0.05）and significantly aggravated the generation of ROS（P〈0.05）.In contrast,the apoptotic ratio was reduced by the application of MDL28170（P〈0.05）.Conclusion：Hyperuricemia can exacerbate the vulnerability and the apoptosis of cardiomyocytes induced by ischemia/hypoxia through the increased generation of ROS via calpains.

关 键 词：高尿酸 钙蛋白酶1 钙蛋白酶2 缺血/缺氧 心肌细胞 

分 类 号：R543.5[医药卫生—心血管疾病]