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0.5～24 h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(Cleaved caspase-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2 h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.
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