基于“肾藏精起亟”理论观察补肾活血复方通过干预klotho改善心梗后心衰大鼠心肌纤维化

Based on the Theory of “Kidney Stores Essence and Urgent” Observing Bushen Huoxue Formula to Improve Myocardial Fibrosis in Rats with Heart Failure after Myocardial Infarction by Intervening Klotho

目的:从"肾藏精起亟"出发观察补肾活血复方对klotho蛋白的干预情况,以及进一步对心梗后心衰大鼠心肌纤维化的影响。方法:选取60只雄性WKY大鼠,从中随机选45只行左冠状动脉前降支结扎术,构造心肌梗死的大鼠模型,再将其随机分为Model组、BSHX组、Captopril组。剩下的15只作为Sham组仅挂线,不结扎。BSHX组给予模型大鼠补肾活血复方(9.2g·kg^(-1)·d^(-1)),Captopril组给予模型大鼠卡托普利(12.5mg·kg^(-1)·d^(-1)),Model组和Sham组同时给予等量的去离子水。用药28d后,使用超声检测并计算左室舒张末期内径(LVDD)、收缩末期内径(LVSD)、短轴缩短分数(FS)和射血分数(EF),免疫印迹法(Western Blot)测定心、肾klotho蛋白及心肌Ⅰ型胶原蛋白(Col 1a1)的水平,HE染色观察心肌细胞纤维化的程度。结果:造模成功并药物干预28d后,与Model比较,BSHX组、Captopril组LVDD、LVSD、FS、EF明显改善(P<0.05)。与Sham组比较,Model组肾、心组织klotho蛋白浓度明显降低,心肌Col 1a1表达上升;与Model组比较,BSHX组、Captopril组肾、心klotho蛋白水平升高,心肌Col 1a1表达下调,以上检验结果差异均有统计学意义(P<0.05)。结论:补肾活血复方(BSHX)可能是通过调控klotho对心梗后心衰大鼠,下调了Col 1a1的表达,达到减轻心肌纤维化的结果,以此来改善心衰。

Objective:To observe the intervention of Bushen Huoxue Formula on klotho protein and further influence on myocardial fibrosis in rats with heart failure after myocardial infarction.Methods:60 male WKY rats were selected,and 45 of them were randomly selected to construct the myocardial infarction model,by ligation of the anterior descending branch of the left coronary artery.And then they were randomly divided into the three groups:Model,BSHX and Captopril.The remaining 15 were used as the Sham group and were only hung up without ligation.The BSHX group was given the Bushen Huoxue Formula (9.2 g/kg/d)for model rats,the Captopril group was given captopril (12.5 mg/kg/d) for the model rats.At the same time,the group Model and Sham were given the same dose of deionized water.After 28 days of medication,we used ultrasound to detect and calculate the left ventricular end-diastolic dimension (LVDD),the left ventricular systolic diameter (LVSD),the fractional shortening (FS)and the ejection fraction (EF),Western blotting to determine the levels of heart and kidney klotho protein and myocardial type I collagen (Col 1a1),and observe the results of myocardial cell fibrosis by HE staining.Results:After successful modeling and drug intervention for 28 days,compared with Model,BSHX,Captopril LVDD,LVSD,FS,and EF were significantly improved (P<0.05).Compared with the Sham group,the concentration of klotho in the heart and kidney tissues of the Model group was significantly reduced,and the expression of Col 1a1 protein increased.Compared with the Model group,the levels of klotho in the heart and kidney tissues of the BSHX group and the Captopril group were increased,and the expression of Col 1a1 protein was down-regulated.The differences in the above test results are statistically significant (P<0.05).Conclusion:The Bushen Huoxue Formula may reduce the expression of Col 1a1 in rats with heart failure after myocardial infarction by regulating klotho to reduce myocardial fibrosis,thereby improving heart failure.