参附注射液对大鼠急性心肌梗死I/R损伤的心肌保护机制研究

Protective mechanism of Shenfu Injection on I/R injury in rats with acute myocardial infarction

目的研究参附注射液对大鼠急性心肌梗死(AMI)再灌注(I/R)损伤的心肌保护作用并探讨其机制。方法选取健康大鼠60只,随机分为空白组、模型组、参附组,各20只。模型组、参附组大鼠采用套扎左冠状动脉前降支以复制AMI模型。造模成功后,模型组给予常规口服阿司匹林肠溶片100 mg、硫酸氢氯吡格雷片75 mg、阿托伐他汀钙片20 mg,每天1次,连续1周;参附组在模型组治疗基础上给予股静脉注射参附注射液(2.1 ml/kg),每日2次,连续1周;空白组大鼠不进行造模处理,给予常规饲养,仅按造模组大鼠用药剂量于每日9时、17时给予腹腔注射等量生理盐水,连续1周。观察各组大鼠7 d内的存活情况,并于末次给药后24 h时处死3组大鼠,观察比较3组大鼠心脏组织病理学变化、心肌组织细胞凋亡指数、lncRNA-GAS5、miRNA-210以及EFNA3的表达及心肌细胞再生程度。结果参附组大鼠的存活率为70.00%,与模型组比较差异有统计学意义(P<0.05)。HE染色可见参附组心肌细胞形态较模型组相比更为接近空白组心肌细胞形态;参附组大鼠的细胞凋亡率、lncRNA-GAS5和EFNA3表达量较模型组明显下降,而miRNA-210相对表达量较模型组明显升高,心肌细胞再生程度显著升高,与模型组比较差异均有统计学意义(均P<0.05)。结论参附注射液在改善急性心肌梗死I/R心肌细胞功能方面具有重要保护作用,为中医药在急性心肌梗死心肌损伤治疗靶点提供思路。

Objective To study the myocardial protective effect of Shenfu Injection on reperfusion(I/R)injury in rats with acute myocardial infarction(AMI)and explore its mechanism.Methods A total of 60 healthy rats were randomly divided into blank group,model group and Shenfu group,with 20 rats in each group.The rats in the model group and Shenfu group were ligated with the anterior descending branch of the left coronary artery to replicate the AMI model.After successful modeling,the model group was given routine oral Aspirin Enteric-coated Tablets 100 mg,Clopidogrel Bisulfate Tablets 75 mg and Atorvastatin Calcium Tablets 20 mg,once a day for 1 week;The Shenfu group was given Shenfu Injection through femoral vein on the basis of the treatment of the model group(2.1 ml/kg),twice a day for 1 week.The rats in the blank group were given routine feeding instead of modeling treatment.Only the rats in the model group were given the same amount of normal saline intraperitoneally at 9:00 and 17:00 every day for 1 week.The survival of rats in each group within 7 days were observed.The rats in the three groups were killed 24 hours after the last administration,and the histopathological changes of heart tissue,cardiomyocyte apoptosis index,expression of lncrna-gas5,mirna-210 and efna3 and degree of cardiomyocyte regeneration in the three groups were observed and compared.Results The survival rate of rats in Shenfu group was 70.00%,higher than model group(P<0.05);HE staining showed that the cardiomyocyte morphology of the Shenfu group was closer to that of the blank group than that of the model group;the apoptosis rate,lncrna-gas 5 and efna 3 expression of the Shenfu group were significantly lower than that of the model group,while the relative expression of mirna-210 was significantly higher than that of the model group,and the degree of cardiomyocyte regeneration was significantly higher than that of the model group statistical significance(All P<0.05).Conclusion Shenfu Injection has an important protective effect on improving I/R myocardial cell function in acute myocardial infarction,which provides ideas for the treatment of myocar dial injury in acute myocardial infarction by Traditional Chinese medicine.