TRIM24通过抑制EndoG核转移保护缺血再灌注诱导的心肌细胞凋亡

摘要

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.%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.