抗纤益心方通过调节RhoA/ROCK2信号通路改善扩张型心肌病大鼠心肌纤维化

Kangxian Yixin Recipe Improving Myocardial Fibrosis in Rats with Dilated Cardiomyopathy Via Regulating RhoA/ROCK2 Signaling Pathway

目的:探讨抗纤益心方抑制扩张型心肌病(dilated cardiomyopathy,DCM)模型大鼠心肌纤维化的相关分子机制。方法:80只Wistar雄性大鼠随机分为正常组(10只)和造模组(70只)。采用呋喃唑酮自饮水法建立DCM大鼠模型,将造模成功大鼠(45只)随机分为模型组、卡托普利(10.125 mg·kg^(-1))组及抗纤益心方高(3.6 g·kg^(-1))、中(1.8 g·kg^(-1))、低剂量(0.9 g·kg^(-1))组。连续药物干预4周后超声心动图检测各组大鼠心脏功能;HE染色和Masson染色检测心肌组织病理改变;ELISA检测血清中可溶性生长刺激表达基因2蛋白(soluble growth stimulating factor 2,sST2)及半乳糖凝集素-3(galectin-3,Gal-3)的含量;Western Blot检测Ras同源蛋白家族成员A(ras homologous family member A,RhoA)、Rho相关卷曲螺旋形成蛋白激酶2(rho associated coiled coil forming protein kinase 2,ROCK2)、肌球蛋白磷酸酶靶标亚基1(myosin phosphatase target subunit 1,MYPT1)、p-MYPT1的蛋白表达水平。结果:与正常组相比,模型组大鼠LVEDD、LVESD显著升高(P<0.01),LVEF、LVFS显著降低(P<0.01),心肌组织胶原蛋白过度沉积,CVF及sST2和Gal-3含量显著升高(P<0.01),心肌组织RhoA、ROCK2、p-MYPT1蛋白表达水平显著升高(P<0.01);与模型组相比,抗纤益心方高、中剂量组和卡托普利组大鼠LVEDD、LVESD显著降低(P<0.05),LVEF、LVFS显著升高(P<0.05),心肌组织病理明显改善,胶原蛋白沉积明显改善,心肌纤维化明显减轻,CVF及血清sST2和Gal-3含量显著降低(P<0.05),心肌组织RhoA、ROCK2、p-MYPT1蛋白表达水平显著降低(P<0.05)。结论:抗纤益心方可能通过下调RhoA/ROCK2信号通路抑制DCM大鼠心肌纤维化的发生。

Objective:To investigate the molecular mechanism of Kangxian Yixin Recipe in inhibiting myocardial fibrosis in dilated cardiomyopathy(DCM)model rats.Methods:Eighty male Wistar rats were randomly divided into control group(10 rats)and model group(70 rats).The DCM rat model was established by furazolidone self-drinking method,and the successfully modeled rats(45 rats)were randomly divided into model group,model group,captopril(10.125 mg·kg^(-1))group,and Kangxian Yixin recipe high(3.6 g·kg^(-1)),medium(1.8 g·kg^(-1)),and low(0.9 g·kg^(-1))dose groups.The cardiac function of the rats in each group was detected by echocardiography after 4 weeks of continuous drug intervention.HE staining and Masson staining was used to detect the pathological changes in myocardial tissue.ELISA was used to detect the content of soluble growth stimulating factor 2(sST2)and galectin-3(Gal-3)in serum.And the protein expression levels of RhoA,ROCK2,MYPT1,and p-MYPT1 were detected by Western Blot.Results:Compared with the control group,the LVEDD,and LVESD of the model group were significantly increased(P<0.01),the LVEF and LVFS were significantly decreased(P<0.01),the myocardial tissue collagen was excessively deposited(P<0.01),and the content of sST2 and Gal-3 was significantly increased(P<0.01),and the protein expression levels of RhoA,ROCK2 and p-MYPT1 in myocardial tissue were significantly increased(P<0.01).Compared with the model group,the LVEDD and LVESD of the Kangxian Yixin recipe high and medium dose groups and the captopril group were significantly decreased(P<0.05),while the LVEF and LVFS were significantly increased(P<0.05).The myocardial tissue pathology was improved,collagen deposition was improved,myocardial fibrosis was significantly reduced,CVF score and serum sST2 and Gal-3 contents were significantly decreased(P<0.05),and the protein expression levels of RhoA,ROCK2,and p-MYPT1 in myocardial tissue were significantly decreased(P<0.05).Conclusion:Kangxian Yixin Recipe could inhibit the occurrence of myocardial fibrosis in DCM rats by down-regulating the RhoA/ROCK2 signaling pathway.