乌头赤石脂丸对心肌缺血再灌注大鼠血管内皮细胞及氧化应激的保护作用  

Protective Effect of Wutou Chishizhi Wan on Vascular Endothelial Cells and Oxidative Stress in Myocardial Ischemia-reperfusion Rats

作　　者：王梦妮 谢璐璐 张兆鹏 凌冰 林致辉 刘派[1] 郭军鹏[1] 刘宏岩[1] WANG Meng-ni;XIE Lu-lu;ZHANG Zhao-peng;LING Bing;LIN Zhi-hui;LIU Pai;GUO Jun-peng;LIU Hong-yan(Changchun University of Chinese Medicine,Changchun 130117,China)

机构地区：[1]长春中医药大学,长春130117

出　　处：《中国实验方剂学杂志》2020年第21期40-47,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：吉林省科技厅科技发展计划项目(20170101007JC)。

摘　　要：目的:探讨乌头赤石脂丸对大鼠心肌缺血再灌注损伤(MIRI)的保护作用,并观察其对大鼠心电图变化、凝血功能、血管内皮细胞及氧化应激等相关机制的影响。方法:40只SD大鼠随机分为正常组、模型组、阳性药物组(尿激酶组)及乌头赤石脂丸组,每组10只,除正常组外,其余均建立大鼠心肌缺血再灌注损伤模型。观察大鼠缺血前10 min,缺血后30min,再灌注30,60,120 min(即T0,T1,T2,T3,T4)后心率(HR)及造模后心电图J点的变化;苏木素-伊红(HE)染色法观察大鼠心肌组织病理形态学变化;观察大鼠凝血4项指标[凝血酶原时间(PT),活化部分凝血活酶时间(APTT),凝血酶时间(TT),纤维蛋白原(FIB)]指标的变化;观察大鼠血清中内皮素-1(ET-1),血栓素A2(TXA2),前列环素(PGI2)含量及心肌组织超氧化物歧化酶(SOD),丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)水平的影响。蛋白免疫印迹法(Western blot)检测氧化应激指标心肌组织蛋白Keap1与核转录因子E2相关因子(Nrf2)表达水平。结果:与正常组比较,MIRI模型组大鼠心电图出现显著心肌缺血损伤样改变,ST段显著抬高,J点显著升高,在T1,T2,T3,T4时期HR显著降低(P<0.05,P<0.01);与模型组比较,乌头赤石脂丸使MIRI大鼠心电图,J点改变显著减轻,在T1,T2,T3,T4时期HR升高(P<0.05,P<0.01);与正常组比较,模型组PT,APTT和TT均显著缩短(P<0.01),FIB含量显著升高(P<0.01);模型组血清中PGI2水平显著降低,TXA2,ET-1含量显著升高(P<0.01),模型组心肌组织SOD含量和GSH-Px活性显著降低(P<0.01),MDA含量升高(P<0.01);与模型组比较,乌头赤石脂丸组PT延长(P<0.05),APTT稍有延长,TT显著延长(P<0.01),FIB含量降低(P<0.05),乌头赤石脂丸组血清PGI2水平升高(P<0.05),TXA2,ET-1含量显著降低(P<0.01),心肌组织MDA含量降低且SOD含量和GSP-Px活性显著升高(P<0.01)。同时乌头赤石脂丸组能够激活Keap1/Nrf2信号通路,使Nrf2表达明显升高,Keap1表达显著下降(P<0.Objective: To investigate the protective effect of Wutou Chishizhi Wan on myocardial ischemia reperfusion injury(MIRI)in rats,and observe its effect on such mechanisms as coagulation function,vascular endothelial cells and oxidative stress in rats. Method:A total of 40 SD rats were randomly divided into normal group,model group,positive drug group(Urokinase group)and Wutou Chishizhi Wan group,with 10 rats in each group. Except for the normal group,rat myocardial ischemia-reperfusion injury models were established. The changes of heart rate(HR)at 10 min before ischemia,30 min after ischemia and 30,60,120 min(T0,T1,T2,T3,T4),and the change of electrocardiogram(ECG)J point after modeling in rats were observed. The pathological changes of rat myocardial tissue were observed by hematoxylin-eosin(HE)staining.The changes of four indexes of coagulation [prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT),fibrinogen content decreased significantly(FIB)]in rats were observed. The contents of endothelin-1(ET-1),thromboxane A2(TXA2)and prostacyclin(PGI2)in serum and myocardium levels of superoxide dismutase(SOD),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)of MIRI rats were observed. Western blot assay was used for the detection of oxidative stress protein Keap1 and transcription factor-E2-related factor(Nrf2)expression levels in rat myocardial tissue. Result: Compared with the normal group,the ECG of MIRI rats showed significant myocardial ischemic injury-like changes,ST segment was significantly elevated,J point was significantly increased,and the incidences of HR in T1,T2,T3 and T4 were significantly reduced(P<0.05,P<0.01). Compared with the model group,Wutou Chishizhi Wan significantly reduced ECG J-point changes in MIRI rats,while increased the incidence of HR in T1,T2,T3 and T4(P<0.05,P<0.01). Compared with the normal group,PT,APTT and TT in the model group were significantly shortened(P<0.01),FIB content was significantly increased(P<0.01),and the serum PGI2 level decreased and

关 键 词：乌头赤石脂丸 心肌缺血再灌注 凝血功能 氧化应激 Keap1/核转录因子E2相关因子(Nrf2) 

分 类 号：R2-0[医药卫生—中医学] R22