低温保存诱导大鼠心肌细胞Smac／DIABLO蛋白表达的动态变化  

DYNAMIC CHANGES OF Smac/DIABLO PROTEIN EXPRESSION IN RAT DONOR MYOCARDIUM SUFFERED FROM HYPOTHERMIC PRESERVATION

作　　者：郑鸣之[1] 蒋建平[1] 傅林[2] 朱立 陈莹莹 沈岳良 

机构地区：[1]浙江医学高等专科学校,浙江杭州310053 [2]浙江大学基础医学国家理科人才培养基地,浙江杭州310058 [3]医学院生理学教研室,浙江杭州310058 [4]浙江大学医学院生理学教研室,浙江杭州310058

出　　处：《中国应用生理学杂志》2009年第3期296-300,I0002,共6页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助(30470635);浙江医学高等专科学校科技计划资助(2008XZA03)

摘　　要：目的:观察大鼠供心不同时程低温保存后线粒体Smac/DIABLO蛋白表达的差异。方法:根据不同的低温保存时程,SD大鼠随机分5组(n=8)。采用Langendorff离体鼠心灌注法停搏大鼠心脏,检测心脏在4℃条件下Cel-sior保存液中分别保存0、3、6、9、12h后,心肌细胞线粒体内超氧化物岐化酶(SOD)活性和丙二醛(MDA)含量的变化。并采用Westorn blotting蛋白印迹分析法观察心肌细胞Smac/DIABLO蛋白表达情况,原位末端标记(TUNEL)染色法检测心肌细胞凋亡。结果:①随着低温保存时间的延长,心肌细胞线粒体内SOD活性随之降低,MDA含量随之升高,心肌细胞凋亡指数也逐渐增高。②随着低温保存时间的延长,Smac/DIABLO蛋白表达逐渐增多,至低温保存6h后最为显著,随后又逐渐减弱。结论:随着低温保存时间的延长,可能由于心肌细胞抗氧自由基的能力逐步减弱,致使诱导细胞凋亡的心肌线粒体Smac/DIABLO蛋白表达逐渐增强,心肌细胞凋亡逐渐增多。Aim： To investigate the effect of different duration of hypothermic preservation on the expression of Smac/DIABLO protein in rat hearts. Methods： The Langendorff model of isolated rat heart was used. After stored in 4℃ Celsior solution for different time （0, 3, 6, 9 or 12 h）, the activity of SOD and the content of MDA in heart mitochondria were measured. Cell apoptosis was detected by TUNEL technique. The expression of Smac/DIABLO protein was also analyzed by Western blotting. Results： ①After hypothermic preservation, the activity of SOD was decreased and the eontent of MDA was increased in rat hearts in a time-dependent manner. ②Prolonged the hypothermie preservation, the percentage of apoptotie cell also enhanced. ③After Long-term of cold preservation, the expression of Smac/DIABLO protein increased at 3-6 h of preservation but decreased after 9 h. Conclusion： Prolonged the hypothermic preservation might lead to the expression of Smae/DIABLO protein and induce cardiomyoeytes apoptosis, which may in turn result in malfunction of cardiomyocytes.

关 键 词：心脏保存 细胞凋亡 Smae/DIABLO 

分 类 号：R331[医药卫生—人体生理学]