慢病毒介导的Sema3A基因转染对心肌细胞凋亡的影响  

Effects of Lentivirus Mediated Sema3A Gene Transfer on Cardiac Myocyte Apoptosis

作　　者：张丽娉[1] 孙彧[1] 陈仁华[1] 邹云增[2] 李毅刚[1] 

机构地区：[1]上海交通大学医学院附属新华医院心内科,上海200092 [2]复旦大学附属中山医院心内科上海市心血管病研究所,上海200032

出　　处：《中国临床医学》2011年第2期133-135,共3页Chinese Journal of Clinical Medicine

基　　金：国家自然科学基金资助项目(编号:30871082;81070154);2010年度上海市优秀学科带头人计划(编号:10XD1402800)

摘　　要：目的：通过慢病毒转染改变心肌细胞Sema3A表达,研究其对原代培养的心肌细胞凋亡的影响。方法：检测0.5-24h缺氧对乳鼠心肌细胞Sema3A的表达的影响。Sema3A慢病毒转染后,蛋白免疫印迹法检测细胞凋亡蛋白酶片段（Cleavedcaspase-3）表达,缺口末端标记法检测凋亡细胞并计算凋亡率。结果：随着缺氧时间延长,Sema3A表达呈下降趋势。增加细胞Sema3A表达,cleaved caspase-3表达上调（P〈0.05）,细胞凋亡率显著增加（P〈0.05）。抑制缺氧2h的心肌细胞Sema3A的表达,cleaved caspase-3表达下调（P〈0.05）,细胞凋亡率下降（P〈0.05）。结论：Sema3A对心肌细胞有诱导凋亡作用。心肌细胞缺氧损伤后,可能通过下调Sema3A表达减少心肌细胞凋亡。Objective：To examine the effects of lentivirus mediated Sema3A gene transfer on cardiac myocyte apoptosis.Methods：Cultured rat neonatal cardiomyocytes were exposed to hypoxic condition for 0.5-24 h,and the expressing of Sema3A was measured.Following the Sema3A gene transfer by lentivirus,the expression of cleaved caspase-3 protein in cardiomyocyte was detected by Western blot,and the changes in cardiomyocyte apoptosis were detected by TUNEL.Results： Hypoxia for 0.5 to 24 h induced significant decrease in expressing of Sema3A in a time-dependent manner.Lentivirus-mediated Sema3A overexpression significantly increased the level of cleaved caspase-3 protein（P〈0.05） and apoptosis（P〈0.05）.Lentivirus-mediated Sema3A inhibition in cardiomyocytes exposed to hypoxic condition for 2h significantly decreased the level of cleaved caspase-3 protein（P〈0.05） and apoptosis（P〈0.05）.Conclusions： Sema3A induced cardiac myocyte apoptosis.Cardiomyocytes may depress the expressing of Sema3A for self-protection from hypoxia-induced apoptosis.

关 键 词：SEMA3A 心肌细胞 慢病毒 凋亡 细胞凋亡蛋白酶片段 

分 类 号：Q343.1[生物学—遗传学]