瓜蒌薤白半夏汤合血府逐瘀汤组方对小型猪痰瘀互结证冠心病模型心肌细胞凋亡及相关蛋白表达的影响  

Effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction on myocardial apoptosis and relevant protein expressions in miniature swine phlegm and blood stasis type coronary heart disease model

作　　者：闫爱国[1,2] 刘建勋[1] 李欣志[1] 董小霞[1] 李宏坤[1] 

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,北京100091 [2]北京洛奇临床检验所检验医学研发中心,北京102206

出　　处：《中国中药杂志》2015年第11期2174-2179,共6页China Journal of Chinese Materia Medica

基　　金：国家“重大新药创制”科技重大专项(2012ZX09301002-004);北京市十病十药项目(Z131100002513006)

摘　　要：传统方药瓜蒌薤白半夏汤合血府逐瘀汤组方在冠心病心肌缺血损伤中对抑制凋亡的研究很少,本实验通过瓜蒌薤白半夏汤合血府逐瘀汤组方对小型猪痰瘀互结证冠心病模型进行干预,观察此组方对该模型心肌细胞凋亡及相关蛋白Bcl-2,Bax,Caspase-3,Caspase-9表达的影响。采用中国实验小型猪15只,随机分成对照组、模型组和痰瘀同治组。模型组和痰瘀同治组以高脂饲料喂养2周后,用介入法行冠状动脉拉伤,然后开始喂药同时继续予高脂饲料喂养8周。对照组以普通饲料喂养10周,不做冠状动脉拉伤。实验结束后处死动物,取梗死交界处心肌,固定,常规制备石蜡切片,采用TUNEL技术检测心肌凋亡,免疫组织化学(immunohistochemical,IHC)技术检测心肌组织的Bcl-2,Bax,Caspase-3,Caspase-9水平。结果显示TUNEL法检测的对照组、模型组及痰瘀同治组的凋亡指数(apoptosis index,AI)分别为0.92%,27.68%,17.28%,痰瘀同治组的细胞凋亡指数明显小于模型组的凋亡指数(P<0.01)。痰瘀同治组与模型组比较,Bcl-2蛋白表达显著增强(P<0.01),Bax蛋白表达显著降低(P<0.01);痰瘀同治组Caspase-3,Caspase-9表达均显著低于模型组(P<0.01)。以上结果表明瓜蒌薤白半夏汤合血府逐瘀汤组方对小型猪痰瘀互结冠心病模型心肌凋亡细胞有明显的保护作用。There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling （TUNEL） assay was used to detect the myocardial apoptosis. The immunohistochemistry （IHC） technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes （AI） for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group （P〈0.01）. Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression （P〈0.01） and lower Bax, Caspase-3 and Caspase-9 protein expressions （P〈0.01）. In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phleg

关 键 词：冠心病痰瘀互结证 瓜蒌薤白半夏汤合血府逐瘀汤 心肌细胞凋亡 TUNEL Bcl-2 Bax Caspase-3 CASPASE-9 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]