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目的:观察丹蒌片对大鼠心肌缺血再灌注损伤(MIRI)的保护作用,并探讨其机制。方法:将80只健康成年雄性Wistar大鼠随机分为假手术组(以0.5%羧甲基纤维素钠溶液10 m L·kg~(-1)灌胃)、模型组(以0.5%羧甲基纤维素钠溶液10m L·kg~(-1)灌胃)、丹蒌片组(0.9 g·kg~(-1))。预防性给药7 d后,结扎大鼠心脏冠脉左前降支,50 min后复灌,制作大鼠心肌缺血再灌注损伤模型。每组取6只大鼠于复灌2 h检测血清乳酸脱氢酶(LDH),肌酸激酶同工酶(CK-MB),内皮素(ET),一氧化氮(NO)水平及心肌组织丙二醛(MDA)及髓过氧化物酶(MPO)水平。剩余大鼠于造模前及复灌48 h进行心脏射血分数检测,每组取6只进行原位末端转移酶标记技术(TUNEL)染色评估心肌细胞凋亡指数,蛋白免疫印迹法(Western blot)检测心肌含半胱氨酸的天冬氨酸蛋白水解酶(Caspase)-3,B细胞淋巴瘤-2(Bcl-2)及Bcl-2相关X蛋白(Bax)蛋白表达。余大鼠行依文思蓝-TTC染色评估心肌梗死面积。结果:复灌2 h时,与模型组比较,丹蒌片组血清LDH及CK-MB均明显降低(P<0.05),血清ET-1明显降低,NO明显升高(P<0.05),心肌组织MDA及MPO明显升高(P<0.05)。复灌48 h时,与模型组比较,丹蒌片组心脏EF值明显上升(P<0.05),心肌梗死范围明显下降(P<0.05),心肌凋亡指数明显下降(P<0.01);与模型组比较,丹蒌片组心肌Caspase-3,Bax蛋白表达明显升高(P<0.05,P<0.01),Bcl-2蛋白表达下降(P<0.05)。结论:丹蒌片在MIRI早期可以保护内皮细胞功能、抑制氧化及炎症浸润,在MIRI晚期可能通过抑制细胞凋亡,缩小心肌梗死面积,改善心功能来实现对心脏的保护作用。
Objective: To observe the protective effect of Dan Kui Tablet on myocardial ischemia-reperfusion injury (MIRI) in rats and its mechanism. Methods: Eighty healthy adult male Wistar rats were randomly divided into sham operation group (intragastric administration of 0.5% sodium carboxymethyl cellulose solution 10 m L · kg -1), model group (0.5% carboxymethylcellulose Sodium Cellulose solution 10m L · kg ~ (-1) intragastrically), danji tablet group (0.9g · kg -1). After prophylactic administration for 7 days, left anterior descending coronary artery was ligated and reperfusion was performed 50 minutes later to establish myocardial ischemia-reperfusion injury model in rats. Six rats in each group were sacrificed for 2 hours to detect the levels of serum LDH, CK-MB, ET, NO and myocardial tissue Malondialdehyde (MDA) and myeloperoxidase (MPO) levels. Left ventricular ejection fraction (LVEF) was measured at 48 h before and after reperfusion in rats. TUNEL staining was used to evaluate the cardiomyocyte apoptosis index in 6 rats in each group. Western blot ) Were used to detect the protein expression of caspase-3, Bcl-2 and Bcl-2 related protein B in myocardium. Myocardial infarction area was assessed by Evans blue-TTC staining in remaining rats. Results: After reperfusion for 2 h, serum LDH and CK-MB were significantly decreased (P <0.05), serum ET-1 significantly decreased, NO significantly increased (P <0.05) Tissue MDA and MPO were significantly increased (P <0.05). Compared with the model group, the EF of heart in Danji tablet group was significantly increased (P <0.05), the range of myocardial infarction was significantly decreased (P <0.05), and the myocardial apoptosis index was significantly decreased (P <0.01) Compared with the model group, the expression of Caspase-3 and Bax protein in Danji tablet group was significantly increased (P <0.05, P <0.01), and Bcl-2 protein expression was decreased (P <0.05). Conclusion: Danjian tablet could protect the function of endothelial cells and inhibit the oxidation and infiltration of inflammatory cells in the early stage of MIRI. In the later period of MIRI, dan-dan tablet may protect heart by inhibiting apoptosis, reducing myocardial infarct size and improving cardiac function.