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背景:研究证实,参附注射液对各种休克、心力衰竭、心肌缺血以及室上性/室性心律失常均有改善治疗作用,对大鼠心肌缺血再灌注损伤具有良好的保护作用。目的:观察参附注射液对急性缺血再灌注损伤大鼠心肌凋亡相关基因Bcl-2,Bax和c-Fos蛋白表达的影响。设计:完全随机分组设计,对照实验。单位:重庆医科大学附属第二医院麻醉科。材料:实验于2004-04/12在重庆医科大学附属第二医院麻醉教研室完成。选用健康成年Wistar大鼠35只,由解放军第三军医大学大坪医院实验动物中心提供。参附注射液是中医“回阳救逆”参附汤的中药配方,其主要成分是人参皂甙及乌头类生物碱(雅安三九药业有限公司出品,规格10mL/支,批号:030110)。方法:采用在体心脏缺血再灌注模型,35只大鼠按随机数字表法分为5组,每组7只。①假手术组:只穿线不结扎,静脉注射生理盐水8mL/kg,观察120min。②参附注射液30min组:结扎前15min静脉注射参附注射液8mL/kg,结扎左冠状动脉前降支40min,再灌注30min。③参附注射液120min组:再灌注120min,余同参附注射液30min组。④生理盐水30min对照组:结扎左冠状动脉前降支前15min静脉注射生理盐水8mL/kg,结扎左冠状动脉前降支40min,再灌注30min。⑤生理盐水120min对照组:再灌注120min,余同生理盐水30min组。应用免疫组化法检测各组大鼠心肌凋亡相关基因Bcl-2,Bax和c-Fos蛋白的表达量。主要观察指标:各组大鼠心肌凋亡相关基因Bcl-2,Bax和c-Fos蛋白的表达量。结果:35只大鼠全部进入结果分析,无脱失。①与假手术组比较,生理盐水30min组和120min组大鼠心肌Bcl-2,Bax和c-Fos蛋白表达水平显著升高(P<0.01),Bcl-2/Bax比率显著降低(P<0.01)。②与相应生理盐水组比较,参附注射液30min组和120minBcl-2蛋白表达水平显著升高(P<0.01),Bax和c-Fos蛋白表达水平显著降低(P<0.01),Bcl-2/Bax比率显著升高(P<0.01)。结论:参附注射液对缺血再灌注心肌保护效应可能与其促进Bcl-2蛋白高表达、抑制Bax与c-Fos蛋白表达、增加Bcl-2/Bax比率,从而抑制心肌细胞凋亡有关。
BACKGROUND: Studies have confirmed that Shenfu injection can improve the treatment of various shocks, heart failure, myocardial ischemia, and supraventricular/ventricular arrhythmias, and has a good protective effect on myocardial ischemia-reperfusion injury in rats. OBJECTIVE: To observe the effect of Shenfu injection on expression of Bcl-2, Bax and c-Fos protein in rats with acute myocardial ischemia reperfusion injury. Design: completely random grouping design, control experiment. Unit: Department of Anesthesiology, the Second Affiliated Hospital of Chongqing Medical University. MATERIALS: The experiment was performed at the Department of Anesthesiology, the Second Affiliated Hospital of Chongqing Medical University from April 2004 to December 12th. 35 healthy adult Wistar rats were selected and provided by the Laboratory Animal Center of Daping Hospital, Third Military Medical University of PLA. Shenfu Injection is a traditional Chinese medicine formula of “Shuiyang Salvage” and Shenfu Decoction. Its main components are ginsenosides and Aconitum alkaloids (produced by Yaan Sanjiu Pharmaceutical Co., Ltd., specifications are 10mL/support, lot number: 030110). . METHODS: In vivo cardiac ischemia-reperfusion model was used. Thirty-five rats were divided into 5 groups according to the random number table method, 7 in each group. 1 sham operation group: only threading without ligation, intravenous saline 8mL/kg, observed 120min. 2 Shenfu injection 30min group: 15min before ligation intravenous Shenfu injection 8mL/kg, ligation left anterior descending coronary artery 40min, reperfusion 30min. 3 Shenfu injection 120min group: reperfusion 120min, the same with the Shenfu injection 30min group. 4 normal saline 30min control group: ligation of the left anterior descending coronary artery 15min before intravenous injection of physiological saline 8mL/kg, ligation of the left anterior descending coronary artery 40min, reperfusion 30min. 5 normal saline 120min control group: reperfusion 120min, with the normal saline 30min group. Immunohistochemistry was used to detect the expression of Bcl-2, Bax and c-Fos proteins related to myocardial apoptosis in each group. MAIN OUTCOME MEASURES: The expression of Bcl-2, Bax and c-Fos proteins related to myocardial apoptosis in each group. Results: All 35 rats were involved in the analysis of the results without any loss. 1Compared with the sham group, the expression of Bcl-2, Bax and c-Fos protein in the myocardial 30 min group and 120 min group was significantly increased (P<0.01), and the Bcl-2/Bax ratio was significantly decreased (P<0.01). ). 2Compared with the corresponding saline group, the expression of Bcl-2 protein in Shenfu injection group was significantly increased at 30 min and 120 min (P<0.01), and the expression of Bax and c-Fos protein was significantly decreased (P<0.01). Bcl-2/ The Bax ratio was significantly increased (P < 0.01). Conclusion: The protective effect of Shenfu injection on myocardial ischemia/reperfusion injury may be related to promoting high expression of Bcl-2 protein, inhibiting Bax and c-Fos protein expression, increasing Bcl-2/Bax ratio, and inhibiting myocardial cell apoptosis.