微管损伤与缺氧心肌细胞线粒体损害的关系研究  

Correlation between microtubule damage and mitochondria damage in hypoxic cardiomyocytes

作　　者：邝勇[1] 黄跃生[1] 

机构地区：[1]第三军医大学西南医院全军烧伤研究所创伤烧伤与复合伤国家重点实验室,重庆400038

出　　处：《第三军医大学学报》2006年第15期1547-1549,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目("973"项目)(2005CB522601);国家自然科学基金资助重点项目(30430680);国家杰出青年科学基金资助项目(30125040)~~

摘　　要：目的探讨心肌细胞在缺氧条件下微管损伤与线粒体损害的关系。方法常规方法分离培养W istar乳鼠心肌细胞,分为正常对照组、缺氧组及缺氧微管解聚组,分别于缺氧30 m in、1 h对3组进行缺氧处理,并对缺氧微管解聚组进行特异性微管解聚剂秋水仙素解聚微管后采用流式细胞仪定量检测α-微管蛋白荧光强度变化,激光共聚焦显微镜检测α-微管蛋白及线粒体形态、分布及荧光强度变化。结果与正常对照组比较,缺氧组α-微管蛋白免疫荧光强度明显减弱(P<0.01);微管断裂,分布紊乱,线粒体分布亦出现紊乱,规律性基本丧失。与缺氧组比较,缺氧并微管解聚组微管断裂、分布紊乱、荧光强度减弱明显(P<0.01);线粒体分布弥散,完全失去规律性,基质变淡显著,荧光染色强度亦明显降低(P<0.01)。结论心肌细胞缺氧早期(30 m in)即出现微管损伤,说明微管损伤可能是导致心肌细胞线粒体损害的因素。Objective To investigate the relationship between microtubule damage and mitochondria damage in hypoxic cardiomyocytes. Methods Cardiomyocytes from neonatal Wistar rat were isolated and cultured for 3 -5 d, then divided into three groups： normal control, hypoxia （cultured in hermetic container containing 1% O2, 5% CO2, 95% N2 ）, hypoxia with microtubule depolymerization （ by adding 4 μmol/L colchicine）. The fluorescence intensity of α-microtubule was determined by flow cytometry, the morph and distribution of α- microtubule and mitochondria were checked by confocal laser scanning microscopy （CLSM） at 30 min, 1 h after the cadiomyocytes being oxygen deficit. Results As compared with the normal control cells, the fluorescence intensity of α- microtubule decreased and the microtubule ruptured and distributed confusedly, and the mitochondria changed into disorder in hypoxic cells. As compared with the hypoxic cells, the distribution of mitochondria confused and the fluorescence intensity decreased in the cells of hypoxia with microtubule depolymerization. Conclusion Microtubule damage occurred at the early stage of hypoxia （30 min）. Microtubule damage may lead to mitochondria damage in hypoxic cardiomyocytes

关 键 词：心肌细胞 缺氧 微管 线粒体 激光共聚焦 流式细胞术 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R329.27[医药卫生—基础医学]