一氧化碳释放分子2通过影响HMGB1分布及表达减轻心肌细胞缺氧损伤

CORM-2 Inhibits Myocardial Hypoxia Injury by Affecting the Distribution and Expression of HMGB1

目的:探讨一氧化碳释放分子(CORM-2)对心肌细胞缺氧的保护作用及其机制。方法:将大鼠心肌细胞H9C2分为4组:正常组(H9C2)、缺氧组(Hypoxia+H9C2)、CORM-2组(CORM-2+Hypoxia+H9C2)和iCORM-2组(iCORM-2+Hypoxia+H9C2)。CCK-8和流式细胞术分别检测心肌细胞活性和凋亡情况;免疫荧光检测HMGB1核浆分布情况;Western Blot检测心肌细胞胞浆内HMGB1表达和总HMGB1的表达情况,以及相关凋亡蛋白表达。结果:缺氧组与正常组比较,细胞活性下降,凋亡率增高(9.5%vs 27.4%,P<0.05),胞浆HMGB1迁移增加,细胞总HMGB1及凋亡相关蛋白表达增加(P<0.05)。CORM-2组与缺氧组相比,细胞的活性升高(P<0.05),凋亡率降低至14.1%,胞浆HMGB1释放量及总表达量减少,Cleaved Caspase-3表达降低,ICORM-2组与缺氧组无明显变化。结论:CORM-2通过影响HMGB1分布及表达减轻心肌细胞缺氧损伤。

Objective：To study the protective effect and mechanism of carbon monoxide-releasing molecule（CORM-2）on hypoxia-induced cardiomyocytes injury.Methods：Rat myocardial cells H9C2 were randomly divided into four groups：H9C2 group（the control group）,Hypoxia＋ H9C2 group,CORM-2＋Hypoxia＋H9C2group and ICORM-2＋Hypoxia＋H9C2 group.Cell viability and apoptosis were detected by CCK-8and flow cytometry,respectively.The distribution of HMGB1 was detected by immunofluorescence.The expression levels of total HMGB1,HMGB1 in cytoplasm and apoptosis protein were measured by Western Blot.Results：Compared with the control group,cell viability in Hypoxia＋H9C2 group was decreased（P〈0.05）,while cell apoptosis was increased（from 9.5%to 27.4%,P〈0.05）.Furthermore,the expression levels of total HMGB1 and apoptosis protein were also increased（P〈0.05）with more migration of HMGB1 from nucleus to cytoplasm.Compared with Hypoxia＋H9C2 group,cell viability in CORM-2 ＋Hypoxia＋ H9C2 group was increased（P〈0.05）,cell apoptosis was decreased to 14.1%,and the expression levels of the total HMGB1 and Cleaved Caspase-3 were inhibited（P〈0.05）,alongwith reduced amount of the migratory HMGB1（P〈0.05）at the same time.The results of ICORM-2＋Hypoxia＋H9C2 group had no significant difference from those of Hypoxia＋H9C2 group.Conclusion：CORM-2 inhibits hypoxia injury of myocardium by affecting distribution and expression of HMGB1.