大黄䗪虫丸对博来霉素致肺纤维化大鼠Rho/ROCK信号通路表达的影响

Effects of Dahuang Zhechong Pills on the expression of Rho/ROCK signal pathway in Bleomycin-induced pulmonary fibrosis in rats

目的:观察大黄䗪虫丸对博来霉素致肺纤维化大鼠Rho/ROCK信号通路表达情况的影响,探讨其防治特发性肺纤维化(IPF)的作用机制。方法:将80只健康雄性SD大鼠随机分为空白对照组(Control组)、模型组(Model组)、吡非尼酮组(PFD组)、大黄䗪虫丸大剂量组(DHZC-H组)、大黄䗪虫丸中剂量组(DHZC-M组)、大黄䗪虫丸小剂量组(DHZC-L组)。采用一次性气管内滴入博来霉素(5 mg/kg)复制肺纤维化大鼠模型;给药28 d后处死大鼠,行ELISA法检测TGF-β1、TNF-α含量;RT-PCR法检测ROCK1、α-SMA mRNA表达;Western blot法检测p-RhoA、ROCK1、p-MLC蛋白表达。结果:与Model组比较,各治疗组TGF-β1浓度和DHZC-L组TNF-α浓度显著降低(P<0.01,P<0.05);DHZC-L组TGF-β1浓度显著低于PFD组(P<0.05)。与Model组和PFD组比较,DHZC-M组和DHZC-L组ROCK1、α-SMA mRNA相对表达量均显著降低(P<0.01,P<0.05)。与Model组比较,大黄䗪虫丸各剂量组p-RhoA蛋白相对表达量以及各治疗组ROCK1、p-MLC蛋白相对表达量显著降低(P<0.05,P<0.01);且DHZC-L组p-RhoA、ROCK1、p-MLC蛋白相对表达量显著低于PFD组(P<0.01)。结论:大黄䗪虫丸可通过下调TGF-β1介导的RhoA/ROCK1信号通路及减少α-SMA mRNA及TNF-α表达,起到抗炎抗纤维化作用。

To observe the effects of Dahuang Zhechong Pills(DHZC)on the expression of Rho/ROCK signaling pathway in bleomycin-induced pulmonary fibrosis rats,and to explore its mechanism of prevention and treatment on idiopathic pulmonary fibrosis(IPF).Methods:Eighty healthy male SD rats were randomly divided into a blank control group(Control group),a model group(Model group),a pirfenidone group(PFD group),a large-dose group of DHZC(DHZC-H group),a medium-dose group of DHZC(DHZC-M group),a small-dose group of DHZC(DHZC-L group).Pulmonary fibrosis rat model was replicated with a one-time instillation of bleomycin(5 mg/kg)into the trachea;the rats were sacrificed 28 d after the administration,the TGF-β1 and TNF-αcontent was detected by the ELISA method;RT-PCR method was used to detect ROCK1 andα-SMA mRNA expression;Western blot method was used to detect p-RhoA,ROCK1 and MLCP protein expression.Results:Compared with Model group,TGF-β1 concentration in treatment groups and TNF-αconcentration in DHZC-L group were significantly decreased(P<0.01,P<0.05);The concentration of TGF-β1 in DHZC-L group was significantly lower than that in PFD group(P<0.05).Compared with Model group and PFD group,the mRNA relative expression levels of ROCK1 andα-SMA in DHZC-M and DHZC-L groups were significantly decreased(P<0.01,P<0.05).Compared with Model group,DHZC every dose group of p-RhoA protein relative expression and each treatment group ROCK1 and p-MLC relative expression protein significantly reduced(P<0.05,P<0.01);The relative expressions of p-RhoA,ROCK1 and p-MLC in DHZC-L group were significantly lower than those in PFD group(P<0.01).Conclusion:DHZC plays an anti-inflammatory and anti-fibrosis effect by down-regulating the RhoA/ROCK1signal pathway mediated by TGF-β1 and reducing the expression ofα-SMA mRNA and TNF-α.