Protective effects of Shenfu Injection on myocardial fibrosis in rats with chronic heart failure based on miR-139/Wnt/β-catenin signaling pathway
Objective To investigate the mechanism of action of Shenfu Injection(SFI)on myocardial fibrosis in rats with chronic heart failure(CHF)induced by isoprenaline(ISO).Methods Forty-five SD rats were divided into normal group(n=9)and modeling group(n=36)by random number table method.CHF rat model was prepared by 14-day subcutaneous multi-point injection of ISO on the back,followed by 14-day normal feeding to validate and evaluate the model.During this period,five rats died and four rats were not modeled.Then the 27 successfully modeled rats were subdivided into three groups by the random number table method:model group(intraperitoneal injection of 6 mL·kg-1 sodium chloride injection+gavage of 10 mL·kg-1 distilled water),SFI group(intraperitoneal injection of 6 mL·kg-1 SFI+gavage of 10 mL·kg-1 distilled water),and captopril group(intraperitoneal injection of 6.0 mL·kg-1 normal saline+gavage of 10 mL·kg-1 distilled water containing 8.8 mg·kg-1 captopril which was clinically equivalent).After 15 d of drug intervention,the related indicators of cardiac function and myocardial fibrosis in each group were examined,including echocardiogram,body mass,heart mass index,and left ventricular mass index.ELISA was used to determine N-terminal pro brain natriuretic peptide(NT-proBNP)content;HE staining and Masson staining were performed to observe the histopathological morphology and fibrosis of the myocardial tissue;RT-qPCR was conducted to examine the mRNA expressions of miR-139,Wnt family member 3a(Wnt3a),β-catenin,glycogen synthase kinase 3β(GSK3β),type I collagen(Col-Ⅰ),type Ⅲ collagen(Col-Ⅲ),α-smooth muscle actin(α-SMA),matrix metalloproteinase-2(MMP-2),and matrix metalloproteinase-9(MMP-9)in the myocardial tissue;Western blot was used to examine the protein expressions of Wnt3a,β-catenin,GSK3β,Col-Ⅰ,Col-Ⅲ,α-SMA,MMP-2,and MMP-9 in it.Results Compared with normal group,the left ventricular ejection fraction(LVEF),left ventricular fraction shortening(LVFS)and body mass of rats in model group decreased(P<0.01),while the left ventricular end diastolic diameter(LVEDD),left ventricular end systolic diameter(LVESD),NT-proBNP content,heart mass index,and left ventricular mass index increased(P<0.01);the miR-139 expression decreased(P<0.01),and the protein and mRNA expressions of Wnt3a,β-catenin,GSK3β,Col-Ⅰ,Col-Ⅲ,α-SMA,MMP-2,and MMP-9 increased(P<0.01);the myocardial tissue showed disordered cell arrangement and significant fibrosis,accompanied by obvious inflammatory infiltration.Compared with model group,the LVEF,LVFS,and body mass of rats in SFI group were higher(P<0.01 or P<0.05),while the LVEDD,LVESD,NT-proBNP content,heart mass index,and left ventricular mass index were lower(P<0.01);the miR-139 expression increased(P<0.01),and the protein and mRNA expressions of Wnt3a,β-catenin,GSK3β,Col-Ⅰ,Col-Ⅲ,α-SMA,MMP-2,and MMP-9 decreased(P<0.01);cell arrangement tended to be neat in the myocardial tissue,with reduced fibrous connective tissue hyperplasia,inflammatory cell infiltration,collagen fiber deposition,and fibrosis degree.Conclusion SFI can improve cardiac function and alleviate myocardial fibrosis in rats with CHF,and its mechanism may be related to the regulation of miR-139/Wnt/β-catenin signaling pathway.
万方数据
维普
