康复新液对转化生长因子-β1诱导人胚肺成纤维细胞MRC-5纤维化模型的干预作用  

Effect of Kangfuxin Liquid in Alleviating Human Fetal Lung Fibroblast MRC-5 Cell Fibrosis Model Induced by Transforming Growth Factor-β1

作　　者：徐秋颖[1] 刘伟伟[1] 王宝家[1] 马秀英 高永翔[1] XU Qiuying;LIU WeiweP;WANG Baojia;MA Xiuying;GAO Yongxiang(Basic Medical College,Chengdu University of Traditional Chinese Medicine,Chengdu 610075,China;Medicinal Periplaneta arrtericana Key I.aboratory of Sichuan Province,Chengdu 610000,China)

机构地区：[1]成都中医药大学基础医学院,四川成都610075 [2]药用美洲大蠊四川省重点实验室,四川成都610000

出　　处：《厦门大学学报（自然科学版）》2018年第3期432-437,共6页Journal of Xiamen University：Natural Science

基　　金：成都市科技惠民项目(2015-HM01-00466-SF)

摘　　要：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:10ng/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.fhen, 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 qRf PCR and Western blotting.The results showed that the pulmonary fibrosis cell model was established by treating human fe tal lung fibroblast MRC 5 cell with TGF-β1 at the close of 10 ng/mI..Kangfuxin liquid could significantly alleviate pulmonary fibrosis degree and efficiently down regulate expression levels of a- SMA, L.N, FN, CTGF, Collagen I and Collagen III in the pulmonary fi brosis cell model,with the comparable effect with Dexamethasone.Thus Kangfuxin liquid could significantly alleviate pulmonary fi brosis degree and might be a potential anti fibrosis drug.

关 键 词：康复新液 肺纤维化 转化生长因子-Β1 MRC-5细胞 

分 类 号：R33[医药卫生—人体生理学]