氯离子通道阻滞剂DIDS和NPPB对缺血再灌注诱导心肌细胞凋亡保护作用的研究  

Protective effect of chloride channel blockers against apoptosis of cardiomyocytes induced by ischemia/reperfusion

作　　者：王晓明[1] 张卫卫[1] 龚卫琴[1] 师堂旺[1] 刘艳[1] 刘佳妮[1] 刘艳霞[1] 

机构地区：[1]第四军医大学西京医院老年病科,西安710032

出　　处：《中华老年心脑血管病杂志》2008年第10期765-767,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家自然科学基金(30570758);陕西省科学技术研究发展计划项目(2005K13-G1-4);西安市科技计划攻关项目(GG05164)资助

摘　　要：目的探讨氯离子通道阻滞剂DIDS和NPPB对缺血再灌注(I/R)诱导心肌细胞凋亡过程中的保护作用及可能机制。方法在原代培养鼠心肌细胞I/R诱导心肌细胞凋亡模型中,将实验分为对照组、I/R组、DIDS组和NPPB组,每组20孔。观察心肌细胞的存活率、caspase-3活性及细胞膜完整性。结果I/R组心肌细胞存活率52.1%,DIDS组和NPPB组分别是84.5%和74.7%,均有统计学差异(P<0.01);细胞凋亡DNA电泳片断显示,DIDS组和NPPB组均能显著的抑制DNA的降解。再灌注24 h后,I/R组细胞内caspase-3活性水平482.3%,DIDS组和NPPB组分别是171.7%和211.8%,均有统计学差异(P<0.01)。各组乳酸脱氢酶水平均<5%。结论在I/R诱导的凋亡性心肌细胞损伤中,氯离子通道阻断滞剂能通过有效的抑制Caspase-3活性,起到保护心肌细胞的作用。Objective To explore the protective effects of chloride channel blockers DIDS and NPPB against apoptosis of cardiomyocytes injured by ischemia/reperfusion（I/R）. Methods Cell viability was measured by MTT assay and lactate dehydrogenase （LDH） release,caspase-3 activity levels were measured by fluorescence spectrophotometry. The DNA fragmentation was examined by electrophoretic analysis. Apoptotic mouse cardiomyocytes model was successfully established by I/R. Mouse cardiomyocytes were used in the experiment and were divided into control group,I/R group, DIDS group and NPPB group. All experiment raw data were analyzed with Kruskal-Wallis and Wilcoxon rank sum tests. Results The cell viability of groups treated with DIDS and NPPB was 84.5% and 74.7% ,respectively;compared with 52.1% in I/R group,there were significant differences （P〈0.01 ）. Degradation of DNA fragment was inhibited in accordance with cell viability. The caspase-3 activity levels of the groups treated with DIDS and NPPB were 171.7% and 211.8% at 24 hours after reperfusion,respectively;compared with 482.3% in I/R group,there were significant differences （P〈0.01）. In all experiment groups, the cell LDH leakage was less than 5%. Conclusion The research proves that chloride channel blockers can protect injured cardiomyocyte by inhibiting caspase-3 activity in ischemia/reperfusion process.

关 键 词：心肌再灌注 细胞凋亡 氯化物通道 心肌缺血 

分 类 号：R541[医药卫生—心血管疾病]