钙蛋白酶抑制剂对大鼠离体心脏缺血/再灌注损伤的保护作用-东南大学学报医学版

钙蛋白酶抑制剂对大鼠离体心脏缺血/再灌注损伤的保护作用

单位：1.东南大学,临床医学院,江苏,南京,210009； 2.东南大学医学院附属扬州医院,心血管病研究所,江苏,扬州,225001

分类号：R541, R-332

出版年·卷·期（页码）：2008·27·第二期（86-90）

摘要：

目的:研究calpain抑制剂(ALLN)对大鼠离体心脏缺血/再灌注损伤的保护作用及其机制.方法:45只雄性SD大鼠随机分为正常灌注组、缺血/再灌注组、ALLN+正常灌注组、ALLN+缺血/再灌注组和二甲亚砜+缺血/再灌注组,分别检测再灌注15 min时冠状动脉流出量(CF)和冠脉流出液中乳酸脱氢酶(LDH)漏出量,荧光分光光度计测定荧光强度来反映calpain活性,免疫印迹法检测心肌细胞calpain蛋白表达量,用流式细胞仪检测细胞内钙荧光强度和细胞凋亡率,并计算凋亡指数.结果:同缺血/再灌注组相比,ALLN+缺血/再灌注组CF显著增高(P<0.01),LDH漏出量、calpain活性和蛋白表达量、胞内钙荧光强度和细胞凋亡指数均显著降低(P<0.01).结论:ALLN对离体心脏缺血/再灌注损伤有保护作用,可能与其抑制calpain活性,从而减轻calpain对细胞的损伤,减轻了细胞内钙超载有关.

Objective To investigate the cardioprotective effects of calpain inhibitor(ALLN) against ischemia-reperfusion(I/R) injury of extracorporeal rat hearts and its mechanism.Methods Forty five male SD rats were randomly separated into 5 groups: control group,I/R group,ALLN+control group,ALLN+I/R group and DMSO+I/R group.Coronary flux(CF)and the value of LDH were detected,respectively.The fluorescence intensity reflecting the activity of calpain was detected by fluorospectrophotometer.The calpain protein expression levels were measured by Western blotting.The fluorescence intensity of intracellular Ca2+and rate of apoptosis were detected by flow cytometer,and then apoptotic indexes were calculated.Results Comparing with the I/R group,in ALLN+I/R group,CF was significantly increased(P<0.01).The value of LDH,the protein content and activity of calpain,the fluorescence intensity of intracellular Ca2+and the index of apoptosis were significantly decreasing(P<0.01).Conclusion ALLN has a significant protective effect on I/R injury of extracorporeal hearts by preventing increase in activity of calpain,which may be related with attenuating cell trauma and lessening the intracellular calcium overload.

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