蛭龙活血通瘀胶囊下调Erbb4-IR通过TGFB1/SMAD3信号途径改善小鼠心肌成纤维细胞纤维化  

Zhilong Huoxue Tongyu(蛭龙活血通瘀)Capsule Down Regulates Erbb4-IR and Inhibits Cardiac Fibroblast Fibrosis in Mice via TGFB1/SMAD3 Signaling Pathway

作　　者：刘孟楠 罗钢 董丽 刘平 刘颜 杨廷富 杨思进 Liu Mengnan;Luo Gang;Dong Li;Liu Ping;Liu Yan;Yang Tingfu;Yang Sji(Department of Cardiovascular Medicine,Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University,Luzhou 646000;Faculty of Chinese Medicine,Macao University of Science and Technology,Macao 999078)

机构地区：[1]西南医科大学附属中医医院心血管内科,泸州1646000 [2]澳门科技大学中医药学院,中国澳门999078

出　　处：《中药药理与临床》2022年第6期70-75,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(编号:82074378);四川省中医药管理局科学技术研究专项课题(编号:2021MS097)。

摘　　要：目的:探讨蛭龙活血通瘀胶囊下调Erbb4IR通过TGFB1/SMAD3信号途径对血管紧张素II(angiotension II,Ang II)诱导的心肌成纤维细胞(cardia cfibroblasts,CFs)纤维化的影响。方法:提取原代CFs,使用免疫荧光法鉴定所得细胞;建立Ang II诱导的CFs纤维化模型,分为空白对照组、正常对照血清组、模型对照组、蛭龙活血通瘀胶囊5%、10%、15%、20%、25%含药血清组、洛沙坦10-5mol/L组,各组分别给予相应药物后MTT法检测CFs的增殖活力;处理细胞后镜下观察细胞形态,用RT-PCR法检测Erbb4ir,Tgfb1、Samd3和Asma mRNA表达;用Western blot法检测TGFB1、p-SMAD3和ASMA蛋白表达;沉默Erbb4ir在CFs模型表达后,用RT-PCR法和Western blot法检测目的RNA和蛋白在CFs中的表达。结果:确定最佳给药浓度后分组为空白对照组、模型对照组、蛭龙活血通瘀胶囊含药血清10%组、洛沙坦10-5mol/L组;与模型对照组比较,各用药组CFs中Erbb4ir、Tgfβ1、Samd3和Asma mRNA表达均显著下调(P<0.01),TGFB1、p-SMAD3和ASMA蛋白表达均显著下调(P<0.05);沉默Erbb4ir在CFs模型表达后,与模型对照组比较,模型+沉默Erbb4ir组Erbb4ir、Tgfb1、Samd3和Asma mRNA表达均显著下调(P<0.01),TGFB1、p-SMAD3和ASMA蛋白表达均明显下调(P<0.05);与模型+沉默Erbb4IR组比较,蛭龙活血通瘀胶囊含药血清10%组和模型+沉默Erbb4IR(si-Erbb4IR)+蛭龙活血通瘀胶囊含药血清10%组各个目的mRNA和蛋白表达比较均无显著统计学意义(P>0.05)。结论:蛭龙活血通瘀胶囊可抑制AngII诱导的CFs纤维化,其机制可能与抑制CFs增殖,下调Erbb4ir,调控Tgfb1、Samd3和Asma的mRNA及蛋白表达有关。Objective:To investigate the effect of Zhilong Huoxue Tongyu(蛭龙活血通瘀)Capsule(ZL)on down regulating Erbb4IR and inhibiting angiotensin Ⅱ(AngⅡ)-induced cardiac fibroblasts(CFs)fibrosis via(transforming growth factor beta 1)TGFB1/sterile alpha motif domain containing 3(SMAD3)signaling pathway.Methods:The original CFs were extracted and identified by immunofluorescence method.The AngⅡ-induced CF fibrosis model was established and divided into control group,blank serum group,model group,ZL containing serum groups(5%,10%,15%,20%,25%)and losartan group(10-5 mol/L).The proliferation activity of CFs was detected by MTT assay after each group was given corresponding drugs.Following the drug treatment,the cell morphology was observed under microscope,and the mRNA expressions of Erbb4ir,Tgfb1,Samd3,and alpha-smooth muscle actin(Asma)were detected by RT PCR.The protein expressions of TGFB1,p-SMAD3 and ASMA were measured by Western blot.Then the expression of Erbb4ir was silenced in CFs model,and RT PCR and Western blot were performed to determine the expression of target RNA and protein in CFs.Results:With the optimal dosage determined,the CFs were divided into control group,model group,ZL containing serum group(10%)and losartan group(10-5 mol/L).Compared with the conditions in model group,the mRNA expressions of Erbb4 ir,Tgfb1,Samd3 and Asma and the protein expressions of TGFB1,p-SMAD3 and ASMA in CFs of each administration group were down regulated(P<0.01,P<0.05).After the silencing of Erbb4ir in the CFs model,compared with the conditions in model group,the mRNA expressions of Erbb4ir,Tgfb1,Samd3 and Asma and the protein expressions of TGFB1,p SMAD3 and ASMA in the model+Erbb4 IR silence group down regulated(P<0.01,P<0.05).Compared with the model+Erbb4 IR silence group,the ZL containing serum group(10%)and model+Erbb4 IR silence+ZL containing serum group(10%)had no significant difference in target mRNA and protein expression(P>0.05).Conclusion:ZL could inhibit AngⅡ-induced CF fibrosis,and the mechanism mi

关 键 词：蛭龙活血通瘀胶囊 心肌纤维化 血管紧张素Ⅱ 转化因子 

分 类 号：R285.5[医药卫生—中药学]