强心复脉颗粒对兔缺血再灌注窦房结细胞凋亡及Bax,Bcl-2,Fas-L蛋白表达的影响

Effect of Qiangxin Fumai granule on myocyte apoptosis of sinus node and Bax,Bcl-2,Fas-L gene protein in rabbits of sinoatrial ischemia/reperfusion

目的:探讨临床有效中药复方强心复脉颗粒防治病窦综合征的分子作用机制。方法:采用兔右冠状动脉根部结扎/放松法制备兔窦房结缺血再灌注模型,运用原位末端标记(TUNEL)法观察强心复脉颗粒对兔右冠状动脉缺血再灌注致窦房结细胞凋亡的影响,并运用免疫组化的方法观察其对细胞凋亡相关基因Bax,Bcl-2,Fas-L蛋白表达的影响,用Imagepro Plus图象分析系统计算Bax,Bcl-2,Fas-L表达平均吸光度值。结果:兔右冠状动脉缺血再灌注所致窦房结损伤可明显诱导兔窦房结细胞发生凋亡,引发Fas-L基因蛋白表达增强,同时显著引发Bax基因蛋白表达增强,Bcl-2/Bax下降;强心复脉颗粒可显著下调Fas-L和Bax基因的蛋白表达,上调Bcl-2/Bax,明显抑制和阻断窦房结细胞发生凋亡。结论:通过调节缺血再灌注后窦房结Bax,Bcl,Fas-L基因的蛋白表达从而抑制和阻断细胞凋亡的发生,可能是中药复方强心复脉颗粒防治病窦综合征窦房结缺血再灌注损伤的分子作用机制。

Objective： To explore the molecular mechanism of Qiangxin Fumai granule（QFG,an effective Chinese composite drug） in preventing and treating sick sinus syndrome（SSS）.Method： Rabbit model of sinoatrial ischemia/reperfusion was established by occluding and loosening the root of right coronary artery.Effect of QFG on cell apoptosis was observed by TUNEL method,and its effect on apoptotic related gene Bax,Bcl-2 and Fas-L gene protein expression was observed by immunohistochemical method.Average light density values of the expression of Bax,Bcl-2 and Fas-L of SAN cells was determined by Imagepro Plus image analysis system.Result： Sinoatrial injury induced by ischemia/reperfusion could cause evident sinoatrial cell apoptosis,enhance Fas-L gene protein expression and obviously enhance Bax gene protein expression,reduce Bcl-2/Bax ratio.QFG could significantly down-regulate Fas-L and Bax gene protein expression,up-regulate Bcl-2/Bax ratio,significantly inhibit and block the sinoatrial cell apoptosis.Conclusion： To inhibit and block the event of cell apoptosis through regulating Bax,Bcl-2 and Fas-L gene protein expression in sinoatrial node after ischemia/reperfusion might be one of the mechanisms of QFG in preventing and treating sinoatrial ischemia/reperfusion might be one of the mechanisms of QFG in preventing and treating sinoatrial isehemia/reperfusion injury of SSS.