痰瘀同治方对心肌缺血再灌注损伤过程中自噬与凋亡相关基因调节作用研究  

Regulatory Effect of Tanyu Tongzhi Recipe on Autophagy and Antiapoptosis Gene of Myocardial Ischemia-reperfusion Injury in Rats

作　　者：唐丹丽[1] 孙明杰[1] 佟琳[2] 崔海峰[1] 隋宇[1] 张华敏[2] 

机构地区：[1]中国中医科学院医学实验中心,北京100700 [2]中国中医科学院中医药信息研究所,北京100700

出　　处：《中国实验方剂学杂志》2016年第4期101-105,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81202630;81373792);中国博士后科学基金特别项目(2014T70201);中国中医科学院自主选题项目(ZZ2014002)

摘　　要：目的:观察痰瘀同治方对心肌缺血再灌注损伤(IR)不同阶段自噬与凋亡相关基因的调控作用。方法:将80只SD大鼠随机分为假手术组、模型组、痰瘀同治方组(43 g·kg^(-1)ig)及3-MA干预组(13.5 mg·kg^(-1)iv)。可逆性冠脉左前降支结扎缺血30 min,再灌注2 h复制心肌缺血(MI)及IR模型,利用全自动生化仪检测血清肌酸激酶(CK),乳酸脱氢酶(LDH)含量;采用RT-PCR法检测各组大鼠心肌自噬基因Beclin-1,心肌微管相关蛋白轻链3蛋白(LC3)及抗凋亡蛋白Bcl-2表达。结果:与假手术组比较,模型组大鼠在缺血及再灌注阶段血清CK,LDH含量及心肌Beclin-1,LC3 mRNA表达均显著增强,且Bcl-2 mRNA表达减弱(P<0.01);与模型组比较,痰瘀同治组能降低缺血期血清CK及再灌注期CK,LDH含量(P<0.05),抑制缺血期Beclin-1 mRNA及再灌注期Beclin-1,LC3 mRNA表达,上调缺血期及再灌注期Bcl-2 mRNA表达(P<0.05,P<0.01);与假手术组,3-MA组比较,痰瘀同治组在缺血期可上调Beclin-1,LC3 mRNA表达(P<0.01)。经相关性检验,缺血期及再灌注期Beclin-1与Bcl-2均呈负相关表达(P<0.01)。结论:痰瘀同治方通过促进缺血期自噬发生及抑制再灌注期自噬过表达,同时促进抗凋亡蛋白Bcl-2表达而发挥其对缺血再灌注损伤心肌的保护作用。Objective： To investigate the regulatory effect of Tanyu Tongzhi （TYTZ） recipe on autophagy and antiapoptosis gene of myocardial ischemia-reperfusion injury in rats. Method： Totally 80 SD rats were randomly divided into sham-operated group, model group, 3-MA intervention （13.5 mg ·kg-1 iv） and TYTZ recipe （43 g·kg-1 ig） groups. The model of ischemia and reperfusion of the myocardium was reproduced by ligation of left descending artery for 30 min and reperfusion for 2 hours in rats. Serum contents of CK, LDH were measured by automatic biochemical analyzer. The expression level of Beclin-1, LC3 and Bcl-2 were determined with RT-PCR. Result： Compared with the sham-operated group, the contents of CK, LDH and the expression of Beclin-1, LC3 mRNA were both significantly increased, while the level of Bcl-2 mRNA were decreased in MI and IR model groups （P 〈 0. 01 ）. Compared with the model group, the contents of CK and LDH in ischemic period and reperfusion period, the expression of Beclin-1 mRNA in ischemic period and Beclin-1, LC3 mRNA in reperfusion period were lower in TYTZ recipe groups, and myocardial Bcl-2 mRNA activities of the groups in ischemic period and reperfusion period were increased （P 〈 0.05, P 〈 0.01 ）. Compared with the sham-operated group and 3-MA group, the expression of Beclin-1, LC3 mRNA in ischemic period were increased in TYTZ recipe groups （P 〈0.01）. Beclin-I expression was found to have negative correlation with Bcl-2 expression of time in ischemia and reperfusion periods by Pearson test （P 〈 0.01 ）. Conclusion： The TYTZ recipe can reduce myocardial ischemia/reperfusion injury by promoting the occurrence of autophagy during ischemia period, inhibiting them in reperfusion and promoting the expression of the anti-apoptotic protein Bcl-2.

关 键 词：心肌缺血再灌注损伤 自噬 凋亡 痰瘀同治方 

分 类 号：R285.5[医药卫生—中药学]