【作者】 杨薇；

【导师】 万福生；

【作者基本信息】 江西医学院 ， 生物化学与分子生物学， 2003， 硕士

【摘要】 目的:建立血管紧张素Ⅱ(AngⅡ)诱导的培养乳鼠心肌细胞凋亡模型，探讨牛磺酸对心肌细胞凋亡的影响及初步机制。方法:实验分为正常对照组、AngⅡ处理组、牛磺酸处理组、维拉帕米处理组及牛磺酸和维拉帕米联合用药组；使用光镜及电镜对凋亡细胞进行形态学观察，通过DNA琼脂糖电泳、末端脱氧核糖核苷酸转移酶(TdT) 介导的dUTP缺口末端标记（TUNEL）及流式细胞仪技术证实凋亡模型，并观察牛磺酸对心肌细胞凋亡的影响；通过免疫组化技术检测分析各组心肌细胞Bcl-2、Fas/FasL、Caspase-3蛋白表达的变化。结果：AngⅡ处理组的心肌细胞表现出典型的凋亡细胞形态学变化；基因组DNA琼脂糖凝胶电泳、TUNEL和流式细胞仪分析证实心肌细胞凋亡的存在，而牛磺酸能明显抑制心肌细胞的凋亡；免疫组化分析显示凋亡组心肌细胞中的Bcl-2蛋白表达下降，Fas/FasL、Caspase-3蛋白水平则升高，而牛磺酸、维拉帕米和联合用药组均能拮抗凋亡细胞中上述蛋白水平的改变（P<0.05）。结论：一定浓度的AngⅡ可以诱导培养乳鼠心肌细胞的凋亡，牛磺酸对这种凋亡有拮抗作用，其机制可能是通过下调凋亡细胞内钙浓度以增加细胞内Bcl-2蛋白的表达，降低Fas/FasL、Caspase-3蛋白表达而实现的。更多还原

【Abstract】 Objection:To set up a model of apoptosis in cultured cardiomyocytes of neonatal rat (NRCs) induced by angiotensin Ⅱ(AngⅡ) and to investigate the effects of taurine(Tau) on it and the expression changes of apoptotic relative genes. Methods:Cultured NRCs were randomly devided into the control group, AngⅡ groups, Tau-treated groups, Verapamil(Ver)-treated group and Tau+Ver-treated group . Apoptotic cardiomyocytes were observed by light microcopy and transmission electron microscope and examined by agarose gel electrophoresis, TUNEL and flow cytometery method .Effects of taurine on the apoptotic myocytes were observed and analysed .The protein expression of Bcl-2, Fas/FasL, Caspase-3 genes were studied with immunohistochemical staining. Results:Cultured NRCs induced by AngⅡ showed typical apoptotic morphological changes. The methods of agarose gel eletrophoresis, TUNEL method and flow cytometery testified the apoptosis of NRCs induced by AngⅡ and the protection of taurine on it. Experiments with immunohistochemical staining showed that AngⅡ decrease Bcl-2 protein expression and increase Fas/FasL, Caspase-3 protein expression（P<0.05）,while taurine blocked the changes of these genes expression. Conclusion: AngⅡ can induce apoptosis of cultured NRCs, while taurine can inhibit it. The mechanism may be responsible for the protein expression changes of Bcl-2 , Fas/FasL and Caspase-3 by Tau inhibitiory action on the Ca2+ influx evoked by AngⅡ.更多还原