机构地区:[1]中国中医科学院西苑医院基础医学研究所,北京市中药药理重点实验室,北京100091
出 处:《中药药理与临床》2020年第3期154-158,共5页Pharmacology and Clinics of Chinese Materia Medica
基 金:国家973项目(编号:2015CB554405);中国中医科学院科技创新团队项目(编号:YS1303);国家科技重大专项(编号:2017ZX09301018)。
摘 要:目的:观察丹红滴丸对缺血再灌注损伤大鼠心肌细胞的保护作用,及其对凋亡通路Bcl-2/Bax-Cyt-C-Caspase-3的影响。方法:SD大鼠随机分为假手术对照组、模型对照组、丹红滴丸(浸膏粉)1.5、3 g生药/kg组。大鼠心脏冠状动脉结扎45 min/松开结扎线3 h的方法建立缺血再灌注损伤模型。心脏冠状动脉结扎即刻十二指肠给药。N-BT染色计算缺血心肌范围;TUNEL法检测细胞凋亡,计算凋亡指数;酶联免疫吸附法检测细胞色素C(Cyt-C)含量;免疫组织化学法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3表达量。结果:与假手术对照组比较,模型对照组心肌梗死范围、心肌细胞凋亡指数显著增高(P<0.01)、心肌组织半胱氨酸蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2基因(Bcl-2)、BCL2-Associated X的(Bax)蛋白表达显著上调(P<0.01),Bcl-2/Bax比值显著下降(P<0.01),血清中Cyt-C含量升高(P<0.01)。与模型对照组比较,丹红滴丸各组大鼠心肌梗死范围明显减小、凋亡指数显著降低、心肌组织Bcl-2蛋白表达上调、Bax蛋白表达下调、Bcl-2/Bax升高(P<0.05或P<0.01);心肌组织Caspase-3蛋白表达显著下调(P<0.01),Cyt-C水平降低(P<0.01)。结论:丹红滴丸对缺血再灌注损伤大鼠心肌具有保护作用,机制与其调节Bcl-2/Bax-Cyt-C-Caspase-3凋亡信号分子从而抑制心肌缺血再灌注损伤大鼠心肌细胞凋亡相关。Objective:To observe the protective effect of Danhong dropping pill on myocardial cells and the effect on Bcl-2/Bax-Cyt-C-Caspase-3 of the apoptosis pathway in rats with ischemia-reperfusion injury.Methods:SD rats were randomly divided into four groups:the sham operation group,the model group,Danhong dropping pill groups(1.5 g/kg and 3 g/kg).The rat model of ischemia-reperfusion injury was established by ligating the coronary artery for 45 min/releasing the ligation line for 3 h.After modeling,Danhong dropping pill was administered into duodenum immediately.Ischemic myocardial area was calculated by N-BT staining.Apoptosis was detected by TUNEL to calculate the apoptosis index.The content of Cyt-C was detected by ELISA.The expressions of Bcl-2,Bax and caspase-3 were measured by immunohistochemistry.Results:Compared with the sham operation group,the myocardial infarct area and apoptosis index were significantly increased in the model group(P<0.01),the expressions of Caspase-3,Bcl-2 and Bax protein were significantly increased(P<0.01),the ratio of Bcl-2/Bax was significantly decreased(P<0.01),and the content of Cyt-C in serum was significantly increased(P<0.01).Compared with the model group,the myocardial infarct area and the apoptosis index in all Danhong groups were decreased,Bcl-2 protein expression in the myocardial tissue was increased and Bax protein expression was significantly reduced,the ratio of Bcl-2/Bax was significantly increased(P<0.05 orP<0.01).The Caspase-3 expressions in the myocardial tissue and Cyt-C levels in serum were significantly reduced in Danhong groups(P<0.01).Conclusion:Danhong dropping pill has the protective effect on myocardial cells of myocardial ischemia reperfusion rats.Its mechanism may be related to regulating Bcl-2/Bax-Cyt-C-Caspase-3 apoptosis signal molecule and inhibiting the myocardial cell apoptosis.
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