Cardiomyocyte death induced by myocardial ischemia and reperfusion - Measurement with recombinant human annexin-V in a mouse model
Cardiomyocyte death induced by myocardial ischemia and reperfusion - Measurement with recombinant human annexin-V in a mouse model
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DOI：
10.1161/01.cir.102.13.1564
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发表时间：
2000-09-26
期刊：
影响因子：
37.8
通讯作者：
Reutelingsperger, CPM
中科院分区：
文献类型：
--
作者：
Dumont, EAWJ;Hofstra, L;Reutelingsperger, CPM
磷脂酰丝氨酸（PS）外化被认为是细胞程序性死亡的最早标志之一。我们研究了使用标记的人重组膜联蛋白-V。一种选择性结合PS的蛋白质，在体内小鼠模型中检测心肌缺血和再灌注（I/R）的心肌细胞死亡。方法和结果-I/R诱导小鼠心脏结扎和随后释放左前降支冠状动脉周围的缝线。在安乐死前30分钟动脉内注射与标记分子融合的膜联蛋白-V（25 mg/kg）。缺血15分钟后排斥融合30分钟，在危险区域中1.4 +/- 1.2%（平均值+/- SD）的心肌细胞为膜联蛋白-V阳性（n=6）。在缺血15分钟后再灌注90分钟（n=7），这增加到11.4 +/- 1.9%，在缺血30分钟后再灌注90分钟（n = 7），这增加到20.2 +/- 3.3%。在对照小鼠中，包括在PS结合位点注射膜联蛋白-V的小鼠，未观察到膜联蛋白-V阳性细胞。DNA凝胶电泳显示，在缺血15分钟后再灌注30分钟后开始出现典型的梯形，表明细胞死亡程序的激活。通过用一种新型的Na+-H+交换抑制剂预处理干预细胞死亡程序，在缺血30分钟后再灌注90分钟的小鼠中，膜联蛋白-V阳性心肌细胞从20.2%显著减少到2.2%。R在心脏和细胞死亡阻断策略的评估。
Introduction-Phosphatidylserine (PS) externalization is regarded as one of the earliest hallmarks of cells undergoing programmed cell death. We studied the use of labeled human recombinant annexin-V. a protein selectively binding to PS, to detect cardiomyocyte death in an in vivo mouse model of cardiac ischemia and reperfusion (I/R).Methods and Results-I/R was induced in mouse hearts by ligation and subsequent release of a suture around the left anterior descending coronary artery. Annexin-V (25 mg/kg) fused to a marker molecule was injected intra-arterially 30 minutes before euthanasia. After 15 minutes of ischemia followed by 30 minutes of repel-fusion, 1.4 +/- 1.2% (mean +/- SD) of the cardiomyocytes in the area at risk were annexin-V positive (n=6). This increased to 11.4 +/- 1.9% after 15 minutes of ischemia followed by 90 minutes of reperfusion (n=7) and to 20.2 +/- 3.3% after 30 minutes of ischemia followed by 90 minutes of reperfusion (n=7). In control mice, including those injected with annexin-V at the binding site of PS, no annexin-V-positive cells were observed. DNA gel electrophoresis showed typical laddering starting after 15 minutes of ischemia followed by 30 minutes of reperfusion, suggesting activation of the cell death program. Intervention in the cell death program by pretreatment with a novel Na+-H+ exchange inhibitor substantially decreased annexin-V-positive cardiomyocytes from 20.2% to 2.2% in mice after 30 minutes of ischemia followed by 90 minutes of reperfusion.Conclusions-These data suggest that labeled annexin-V is useful for in situ detection of cell death in an in vivo model of I/R in the heart and for the evaluation of cell death-blocking strategies.