三参饮对病毒性心肌炎慢性期TGFβ-Smad通路的干预研究

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目的观察TGFβ-Smad传导通路在病毒性心肌炎慢性期心肌纤维化形成中的作用及三参饮治疗机制。方法 120只健康雄性Balb/c小鼠随机分组,其中30只采用间断多次腹腔注射柯萨奇病毒B3的方法,建立VMC心肌纤维化模型,另外10只注射不含病毒的Eagle’sMEM液作为正常对照组。两个月后模型制作成功。存活的小鼠随机随机分为5组,模型组、卡托普利组、三参饮高、中、低剂量组。分别给予卡托普利及不同浓度三参饮进行治疗,每日灌胃给药1次,45 d后结束。采用ELISA检测血清Ⅰ型、Ⅲ型胶原,采用半定量RT-PCR法检测转化生长因子(TGFβ1)的基因表达。免疫组化染色检测小鼠心肌Smad2/3,Smad7蛋白表达情况。结果感染CVB3后,ELISA检测模型组血清Ⅰ型、Ⅲ型胶原明显增高。TGFβ1表达水平较正常组升高,Smad2/3蛋白表达也升高,而smad7表达减少,与正常组相比差异均有统计学意义(P<0.05)。经过不同浓度三参饮及卡托普利治疗后,血清Ⅰ型、Ⅲ型胶原表达明显减少,TGFβ1、Smad2/3表达均有下降,而Smad7表达升高,差异均有统计学意义(P<0.05),与正常组相比差异有统计学意义(P<0.05)。结论阻断TGFβ-Smad通路可能是三参饮减少心肌组织Ⅰ、Ⅲ型胶原表达以抑制慢性病毒性心肌炎心肌纤维化的机制之一。 Objective To investigate the role of TGFβ-Smad pathway in the pathogenesis of chronic myocardium fibrosis in viral myocarditis and its mechanism. Methods A total of 120 healthy male Balb / c mice were randomly divided into 3 groups. 30 of them were injected intraperitoneally with Coxsackievirus B3 intraperitoneally to establish model of myocardial fibrosis in VMC. The other 10 mice were injected with Eagle’s SMEM solution without virus Normal control group. Two months later the model was made. The surviving mice were randomly divided into 5 groups: model group, captopril group, three-drink high, medium and low dose groups. The patients were given captopril and different concentrations of Shiji ginseng for treatment. The rats were given gavage once daily for 45 days. Serum type I and type III collagen were detected by ELISA, and the gene expression of transforming growth factor (TGFβ1) was detected by semi-quantitative RT-PCR. Immunohistochemical staining was used to detect the expression of Smad2 / 3 and Smad7 protein in myocardium of mice. Results After CVB3 infection, the serum levels of type Ⅰ and type Ⅲ collagen in the model group were significantly increased. Compared with the normal group, the expression of TGFβ1 increased, the expression of Smad2 / 3 protein increased, while the expression of smad7 decreased compared with the normal group (P <0.05). The serum levels of type Ⅰ and type Ⅲ collagen decreased significantly after treated with different concentrations of three herbs and captopril, the expressions of TGFβ1 and Smad2 / 3 decreased, while the expressions of Smad7 increased (P < 0.05), compared with the normal group, the difference was statistically significant (P <0.05). Conclusion Blocking the TGFβ-Smad pathway may be one of the mechanisms by which San Shen Yin can reduce the expression of type I and type III collagen in myocardium to inhibit myocardial fibrosis in chronic viral myocarditis.
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