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目的:观察黄芪甲苷联合羟基红花黄色素A预处理对心肌梗死小鼠心梗面积、心肌细胞凋亡及炎症因子表达的影响。方法:50只(SPF级)C57小鼠雌雄各半,随机分为假手术组(Sham),心肌梗死模型组(MI),黄芪甲苷预处理组(MI+AsⅣ,3.5 mg·kg-1),羟基红花黄色素A预处理组(MI+HSYA,10 mg·kg-1),黄芪甲苷+羟基红花黄色素A联合预处理组(MI+AsⅣ+HSYA,3.5 mg·kg-1+10 mg·kg-1)。造模前连续腹腔注射7 d,每天1次;结扎左冠状动脉前降支建立小鼠心肌梗死模型,用伊文思蓝-TTC双染色方法区分心肌梗死后心肌的梗死区、缺血危险区(AAR)和左心室总面积(LV);Image J软件计算心肌梗死面积,蛋白免疫印迹法(Western blot)检测心肌细胞中B细胞淋巴癌/白血病-2基因(Bcl-2),Bcl-2相关X蛋白(Bax),活化的天冬氨酸蛋白水解酶-3(cleaved-Caspase-3)的表达水平,酶联免疫吸附法(ELASA)检测血清白细胞介素-6(interleukin-6,IL-6),肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的含量。结果:与模型组相比,黄芪甲苷预处理组、羟基红花黄色素A预处理组及联合预处理组均可显著减小小鼠心肌梗死面积,且联合预处理组效果优于单药预处理组(P<0.05,P<0.01);与假手术组相比,模型组的缺血面积和梗死面积增大,单独给药组与联合给药组的缺血面积及梗死面积均减小(P<0.05,P<0.01);与模型组与假手术相比,黄芪甲苷联合羟基红花黄色素A可上调心肌细胞中抗凋亡蛋白Bcl-2的表达,下调促凋亡蛋白cleaved-Caspase-3和Bax的表达;黄芪甲苷与羟基红花黄色素A联合可降低心肌梗死小鼠血清中IL-6,TNF-α的含量(P<0.05,P<0.01)。结论:黄芪甲苷联合羟基红花黄色素A可减小小鼠心肌梗死面积,抑制心肌梗死后的心肌细胞凋亡及炎症反应,发挥保护心肌损伤的作用。
Objective: To observe the effects of Astragaloside combined with hydroxysafflor yellow A pretreatment on the myocardial infarction area, myocardial cell apoptosis and the expression of inflammatory cytokines in myocardial infarction mice. Methods: Fifty (SPF) C57 mice were randomly divided into sham operation group, MI group, MI + AsⅣ (3.5 mg · kg-1) (MI + AsⅣ + HSYA, 3.5 mg · kg -1), pretreatment with hydroxysafflor yellow A (MI + HSYA, 10 mg · kg -1) and astragaloside plus hydroxysafflor yellow A 1 + 10 mg · kg-1). The model of myocardial infarction was established by ligation of the anterior descending artery of the left anterior descending artery. The infarcted area of the myocardium after myocardial infarction and the ischemic risk area ( (AAR) and total left ventricular area (LV). Image J software was used to calculate the area of myocardial infarction. Western blot was used to detect Bcl-2 and Bcl-2 in cardiomyocytes X, Bax, cleaved-Caspase-3, and the levels of serum interleukin-6 (IL- 6), tumor necrosis factor-α (TNF-α) content. Results: Compared with the model group, the pretreatment with astragaloside, the pretreatment with hydroxysafflor yellow A and the combined pretreatment significantly reduced the area of myocardial infarction in mice, and the combined pretreatment was superior to monotherapy Pretreatment group (P <0.05, P <0.01). Compared with the sham operation group, the ischemia area and the infarction area of the model group increased. The ischemia area and the infarct area of the model group and the combination group decreased (P <0.05, P <0.01). Compared with the model group and sham operation, Astragaloside combined with hydroxysafflor yellow A could up-regulate the expression of anti-apoptotic protein Bcl-2 in cardiomyocytes and downregulate the pro-apoptotic protein cleaved-Caspase-3 and Bax. Astragaloside combined with hydroxysafflor yellow A could decrease the levels of IL-6 and TNF-α in the serum of myocardial infarction mice (P <0.05, P <0.01). Conclusion: Astragaloside combined with hydroxysafflor yellow A can reduce the area of myocardial infarction in mice, inhibit myocardial cell apoptosis and inflammatory response after myocardial infarction, and play a role in protecting myocardial injury.