Influence Of Hirudin On The TGF-?1/Smads Signal Pathway Transformingof Human Fetal Lung Fibroblasts MRC-5 Which Was Stimulated By TGF-?1

Objective:By study the influence of Hirudin on human fetal lung fibroblasts MRC-5 of TGF-?1/Smads signal pathway transforming which was stimulated by transforming growth factor-beta 1(TGF-?1),we explore whether Hirudin could effect Lung fibroblast(LFB)that was activated by TGF-?1 or not,in order to provide experimental evidence for the treatment of Pulmonary fibrosis(PF)by traditional Chinese medicine.Methods:We cultured MRC-5 cells in vitro and hirudin as the experimental drug.We divided the MRC-5 cells into five groups randomly,including control group,TGF-?1 group(induced 24 hours by TGF-?1 10ng/ml),Hirudin low concentration group A(induced 24 hours by TGF-?1 10ng/ml later,inject Hirudin 0.1562mmol/L),Hirudin middle concentration group B(induced 24 hours by TGF-?1 10ng/ml later,inject Hirudin 0.3125mmol/L),Hirudin high concentration group(induced 24 hours by TGF-?1 10ng/ml later,inject Hirudin 0.625mmol/L).CCK-8 method was used to detects cell proliferation;TGF-? R?,?-SMA,Smad 3,Smad 7 m RNA and protein were detected by RT-PCR and Western Blotting.The content of Col-?,Col-? were detected by immunofluorescence method.Results:Hirudin can inhibit the proliferation of MRC-5.Compared with the control group,TGF-?1 group can significantly increase the expre-ssion of TGF-? R?,?-SMA,Smad 3 m RNA and protein,and reduce the expression of Smad 7 m RNA and protein,increase the secretion of Col-?,Col-?;Compared with TGF-?1 group,all hirudin groups can reduce the level of TGF-? R?,?-SMA,Smad 3 m RNA and protein,and increase the level of Smad 7 m RNA and protein,reduce the secretion of Col-? and Col-?.Conclusion:Hirudin can inhibit the proliferation and collagen synthesis of MRC-5 that was induced by TGF-?1,regulate signal pathway of TGF-?1/Smads,so as to inhibit the TGF-?1 activation on LFB,and to play the role of anti-pulmonary fibrosis.It is achieved by inhibiting TGF-? R?,?-SMA,Smad 3 m RNA and protein level,increase the level of Smad 7 m RNA and protein,reduce the secretion of Col-? and Col-?.