基于PDGF通过RhoA/ROCK/HIF1α信号通路研究参竹心康方抗心肌纤维化作用机制OA

Objective To investigate whether Shenzhu Xinkang Formula(参竹心康方,SZXKF)can ameliorate isoproterenol(ISO)-induced myocardial fibrosis(MF)in rats by regulating the platelet-derived growth factor(PDGF)-mediated Ras homolog gene family member A(RhoA)/Rho-associated coiled-coil kinase(ROCK)/hypoxia-inducible factor 1α(HIF1α)signaling pathway.Methods An MF rat model was established by subcutaneous injection of ISO.The MF model rats were randomly divided into the Control group,Model group,Enalapril maleate(Enalapril)group,and SZXKF(SZXK)group,with treatments administered for 4 weeks.Hematoxylin-eosin(HE)staining was used to observe histopathological changes in myocardial tissue.Masson's trichrome staining was performed to evaluate collagen fiber deposition in myocardial tissue.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of Collagen I and Collagen III in myocardial tissue.Quantitative real-time reverse transcription polymerase chain reaction(qRT-PCR)was employed to detect the mRNA expression of PDGFRα in myocardial tissue.Western blot(WB)was conducted to assess the protein levels of α-smooth muscle actin(α-SMA),RhoA,ROCK1,ROCK2,and HIF1αin myocardial tissue.Results 1)Pathological staining results(HE and Masson):Compared to the Control group,myocardial cells in the Model group exhibited edema,significant enlargement in volume,and disordered arrangement,with markedly increased collagen deposition in the myocardial intersti-tial space.In contrast,both the Enalapril and SZXK groups showed significant improvement in cardiac pathological morphology com-pared to the Model group.2)ELISA results:Collagen I and Collagen III levels in myocardial tissue were significantly elevated in the Model group versus the Control group(P<0.05).Compared to the Model group,both the Enalapril and SZXK groups demonstrated sig-nificant reductions in Collagen I and Collagen III levels(P<0.05).3)qRT-PCR results:PDGFRαmRNA expression was significantly upregulated in the Model group compared to the Control group(P<0.05).In contrast,PDGFRαmRNA expression was markedly down-regulated in both the Enalapril and SZXK groups compared to the Model group(P<0.05).4)WB analysis:Protein expression levels of α-SMA,RhoA,ROCK1,ROCK2,and HIF1α were significantly increased in the Model group versus the Control group(P<0.05).Compared to the Model group,the Enalapril group showed significant reductions in RhoA,ROCK1,ROCK2,and HIF1α protein expression(P<0.05),but no significant difference in α-SMA expression(P>0.05).The SZXK group exhibited significant decreases in α-SMA,RhoA,ROCK1,ROCK2,and HIF1α protein levels(P<0.05).Conclusion SZXKF exerts anti-MF effects by downregulating PDGF to inhibit the RhoA/ROCK/HIF1α signaling pathway,thereby reducing excessive collagen fiber deposition in myocardial tissue of fibrotic rats.