温阳益心中药预处理对缺血大鼠心肌细胞凋亡的实验研究

Experimental Study of TCM Preconditioning of Warming Yang and Supplementing Heart Qi on Cardiomyocytes Apoptosis of Myocardial Ischemic Rats

目的观察温阳益心中药预处理和心肌缺血预处理对缺血再灌注48h缺血大鼠心肌Bcl-2、Bax、细胞凋亡率的影响。方法选用雄性Wistar大鼠,随机分为4组,其中3组正常喂食,1组灌胃中药悬液,10d后选各组健康大鼠行冠脉结扎,建立大鼠心肌缺血模型。假手术组:完成全部手术操作,只穿线不结扎冠脉;心肌缺血-再灌注组(IR组):模型建立后,心肌缺血45min后再灌注48h;温阳益心中药预处理缺血-再灌注组(中药组):模型建立后,心肌缺血45min后再灌注48h;预适应处理缺血-再灌注组(IP组):模型建立后,心肌缺血5min,再灌注5min,重复3次,之后心肌缺血45min,再灌注48h。采用免疫组化检测Bcl-2、Bax阳性细胞表达,原位杂交检测Bcl-2、BaxmRNA阳性表达;流式细胞仪测定细胞凋亡率(AP)。结果 IP组和中药组AP较IR组低,Bcl-2阳性表达心肌细胞数与Bcl-2mRNA表达率比IP组高,Bax阳性表达心肌细胞数与BaxmRNA表达率比IR组低(P<0.05)。结论 IP与温阳益心中药对IR的心肌保护均具有抑制心肌细胞凋亡的功能,且有等效性。

Objective To observe the effects on the Bcl-2, Bax and rate of cardiomyocytes apoptosis oI rats with ischemia reperfusion for 48 hours treated by preconditioning of warming yang and supplementing heart qi and myocardial ischemic preconditioning. Methods Male Wistar rats were randomly divided into 4 groups, three groups of which were fed normally, one of which was administered with suspension of Chinese medicine. 10 days later, normal rats in each group were operated by coronary artery ligation to establish myocardial ischemia model. The sham-operated group were performed in the same way as myocardial ischemia rats except the coronary artery was not ligated. The ischemia reperfusion （IR） group were reperfused for 24 hours after 45 rain of myocardial ischernia. The TCM group were treated with suspension of Chinese medicine administration of 0.36 g/100 g for 10 days before the same ischemia and reperfusion as IR group. And the myocardial ischemic preconditioning （IP） group were also performed the same ischemia and reperfusion as IR group after 5 min of ischemia and 5 rain of repelfusion for three times. The positive ceils expression of Bcl-2 and Bax was observed by immunohistochemistry. The positive gene expression of Bcl-2 and Bax was detected by hybridization in situ. Cardiomyocytes apoptosis rate was determinated by flow cytometry. Results CeUs apoptosis rate of IP Chinese medicine was lower than IR group. The positive ceils expression of Bcl-2 and Bcl-2mRNA was more significantly than IP group, but was lower than IR group （P 〈0.05）. Conclusion Myocardial ischemic preconditioning and Chinese medicine group of warming yang and supplementing heart qi have the similar effects on protecting cardiomyocytes by myocardial ischemic preconditioning, through inhibiting cardiomyocytes apoptosis.