摘要
目的探究TRIM24在缺血再灌注诱导的心肌细胞凋亡中的作用及分子机制。方法建立H9C2心肌细胞缺血再灌注损伤模型。首先用流式细胞术检测凋亡情况,并用Western blot检测缺血再灌注诱导的心肌细胞凋亡中TRIM24以及凋亡相关蛋白的表达情况。随后通过表达TRIM24验证其抗凋亡作用及分子机制,利用细胞核浆蛋白分离试剂盒提取细胞核和细胞质蛋白,并检测EndoG的核浆分布情况。结果首先确定缺血再灌注损伤能明显诱导心肌细胞凋亡,而且TRIM24在凋亡心肌细胞中的表达明显减少;过表达TRIM24通过调控BAX/BCL-2表达改善线粒体稳态,进而减少线粒体内凋亡相关蛋白EndoG的核转移,最终显著缓解缺血再灌注诱导的心肌细胞凋亡。结论 TRIM24通过调控BAX/BCL-2表达改善线粒体稳态,进而减少线粒体内凋亡相关蛋白EndoG的核转移,最终显著缓解缺血再灌注诱导的心肌细胞凋亡。
Objective To explore the role and molecular mechanism of TRIM24 in ischemia-reperfusion-induced cardiomyocyte apoptosis.Methods A model of H9C2 cardiomyocyte ischemia-reperfusion injury was established.Apoptosis was detected by flow cytometry,and the expression of TRIM24 and apoptosis-related proteins in ischemia-reperfusion-induced cardiomyocyte apoptosis was detected by Western blot.Subsequently,TRIM24 was used to verify its anti-apoptotic effect and molecular mechanism,and the nuclear and cytoplasmic proteins were extracted by nucleocapsid protein isolation kit,and the distribution of EndoG nucleoplasm was detected.Results Ischemia-reperfusion injury induced significantly cardiomyocytes apoptosis,and the expression of TRIM24 was decreased in apoptotic cardiomyocytes.Overexpression of TRIM24 ameliorated mitochondrial homeostasis by regulating the expression of BAX and BCL-2,which could reduce the nucleus translocation of apoptosis-associated protein EndoG from mitochondria.Finally,TRIM24 overexpression significantly reduced cardiomyocytes apoptosis induced by ischemia-reperfusion injury.Conclusion TRIM24 ameliorates mitochondrial homeostasis by regulating the expression of BAX and BCL-2,which could reduce the nucleus translocation of apoptosis-associated protein EndoG from mitochondria,and ultimately reducing cardiomyocytes apoptosis induced by ischemia-reperfusion injury.
作者
王沙
王惠君
黄晓燕
王宗超
宋宝国
吴娟
Wang Sha;Wang Huijun;Huang Xiaoyan;Wang Zongchao;Song Baoguo;Wu Juan (Department of Cardiovascular Surgery
出处
《中国循证心血管医学杂志》
2019年第1期55-58,共4页
Chinese Journal of Evidence-Based Cardiovascular Medicine
基金
2017年陕西重点研发计划项目(2017SF-093)
