解毒通络方对异丙肾上腺素诱导大鼠心肌纤维化作用及机制研究

Effect and Mechnism of Jiedu Tongluo Formula on Myocardial Fibrosis Induced by Isoproterenol in Rats

目的基于探讨糜酶/血管紧张素(Ang)Ⅱ途径探讨解毒通络法治疗心肌纤维化的机制。方法40只Wistar大鼠随机分为正常对照(Ctrl)组、异丙肾上腺素模型(ISO)组、解毒通络方[灌胃解毒通络方50 g/(kg·d),JTF]组和卡托普利治疗[卡托普利0.005 g/(kg·d),CAP]组,每组10只。各组行HE染色及Masson染色评价心肌纤维化程度;碱水解法测定各组大鼠羟脯氨酸含量;ELISA法测定各组心肌组织AngⅡ含量;Western Blot及相对定量实时PCR分别检测各组大鼠心肌组织糜酶蛋白及mRNA表达量。结果与Ctrl组比较,ISO组大鼠心肌组织胶原纤维面积、羟脯氨酸和AngⅡ含量、糜酶蛋白和mRNA表达增加(P<0.05,P<0.01)。与ISO组比较,JTF及CAP组大鼠心肌组织胶原纤维面积、羟脯氨酸和AngⅡ含量、糜酶蛋白mRNA表达下降(P<0.05,P<0.01),JTF组大鼠心肌组织糜酶蛋白含量下降(P<0.05)。与CAP组比较,JTF组大鼠心肌组织糜酶蛋白mRNA表达降低(P<0.05)。结论解毒通络方能够抑制和延缓大鼠心肌纤维化进程,其机制与抑制糜酶/AngⅡ途径有关。

Objective To investigate the effect and mechnism of Jiedu Tongluo Formula(JTF)on myocardial fibrosis based on chyme/angiotensinⅡ(AngⅡ)pathway.Methods Totally 40 rats were randomly divided into control group(Ctrl),isoproterenol model group(ISO),JTF group(50 g/kg per day)and captopril group(CAP,0.005 g/kg per day),10 in each group.HE staining and Masson staining were used to evaluate myocardial fibrosis.Alkaline hydrolysis method was used to detected hydroxyproline content.ELISA method was used to test AngⅡcontent in cardiac tissue.Western Blot and quantitative Real-time PCR were used to detect protein and mRNA expression of chymase in cardiac tissue respectively.Results Compared with Ctrl group,collagen fiber area of myocardial,content of hydroxyproline and AngⅡ,protein and mRNA expression of chymase increased in ISO group(P<0.05,P<0.01).Compared with ISO group collagen fiber area of myocardial,content of hydroxyproline and AngⅡ,mRNA expression of chymase decreased in JTF and CAP group(P<0.05,P<0.01),protein expression of chymase decreased in JTF group(P<0.05).Compared with CAP group,mRNA expression of chymase decreased in JTF group(P<0.05).Conclusion JTF could inhibit and delay the progression of myocardial fibrosis in rats,the mechnism may related to inhibit chymase/AngⅡpathway.