为探讨康复新液对人胚肺成纤维细胞纤维化模型的干预作用,以转化生长因子(transforming growth factor,TGF)-β1诱导人胚肺成纤维细胞MRC-5构建细胞纤维化模型,并运用CCK-8(cell counting kit-8)法检测康复新液对MRC-5细胞的毒性.以测得的最高不致死给药浓度(80倍稀释)的康复新液干预TGF-β1诱导的MRC-5细胞模型,进而通过实时荧光定量PCR(qRT-PCR)和蛋白质免疫印迹(WB)检测细胞纤维化指标的mRNA及蛋白质表达水平变化.结果显示:10 ng/mL TGF-β1可成功诱导MRC-5细胞发生纤维化,且细胞内α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、层黏连蛋白(laminin,LN)、纤维连结蛋白(fibronectin,FN)、结缔组织生长因子(connective tissue growth factor,CTGF)、Ⅰ型胶原蛋白(CollagenⅠ)和Ⅲ型胶原蛋白(CollagenⅢ)的mRNA表达水平均显著升高; 以80倍稀释的康复新液为最高不致死给药浓度,对MRC-5细胞纤维化模型进行干预,上述纤维化指标的mRNA及蛋白质表达水平均出现显著下降,与地塞米松(Dexamethasone,DXM)的干预效果相当.综上,康复新液可改善由TGF-β1诱导MRC-5细胞纤维化的表型,有望被应用于肺纤维化的治疗.
To investigate the effect of Kangfuxin liquid on the pulmonary fibrosis cell model,the pulmonary fibrosis cell model was established by treating human fetal lung fibroblast MRC-5 cell with transforming growth factor-β1(TGF-β1),and the toxic effect of Kangfuxin liquid on MRC-5 cell was detected by cell-counting kit-8(CCK-8)assay.Then,Kangfuxin liquid diluted by 80 times was implemented on MRC-5 cell model induced by TGF-β1,and the mRNA and protein expression levels of fibrosis indexes were detected by qRT-PCR and Western blotting.The results showed that the pulmonary fibrosis cell model was established by treating human fetal lung fibroblast MRC-5 cell with TGF-β1 at the dose of 10 ng/mL.Kangfuxin liquid could significantly alleviate pulmonary fibrosis degree and efficiently down-regulate expression levels of α-SMA,LN,FN,CTGF,Collagen Ⅰ and Collagen Ⅲ in the pulmonary fibrosis cell model,with the comparable effect with Dexamethasone.Thus Kangfuxin liquid could significantly alleviate pulmonary fibrosis degree and might be a potential anti-fibrosis drug.
