多西环素对TGF-β1诱导肺成纤维细胞α-SMA、FN、Col Ⅰ表达的影响

目的 研究多西环素(Doxy)对转化生长因子-β1 (TGF-β1)诱导人胚肺成纤维细胞(MRC-5)向肌成纤维细胞转化的影响,探索Doxy抗肺纤维化的作用.方法 体外培养MRC-5,采用不同浓度的Doxy(0、2.5、5.0、10.0、20.0、40.0、80.0 μg/mL)干预,72 h后采用MTT法检测细胞活性...

Effect of doxycycline on the expression of alpha-SMA,fibronectin and collagen I in MRC-5 cell line induced by TGF-beta1

Objective To explore the effect of doxycycline(Doxy)on Transforming growth factor-β1(TGF-β1)induced conversion in lung fibroblast,and to provide a theoretical basis for the treatment of pulmonary fibrosis.Methods Human embryonic lung fibroblast(MRC-5)were cultured in vitro,treated with different concentrations of doxycycline(0,2.5,5.0,10.0,20.0,40.0,80.0μg/mL)for 72 hours,3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to detect the cell survival to determine the concentration of doxycycline.TGF-β1 was used to induce the conversion in MRC-5 cell,and treated with different concentrations of doxycycline(0,2.5,5.0,10.0,20.0μg/mL)for 72 hours.Western blotting,RT-PCR,immunity fluorescence were used to detect the expression ofα-smooth muscle actin(α-SMA),fibronectin(FN),collagenⅠ(ColⅠ).Results The MRC-5 cell viability decreased which the concentration of Doxy was 40.0μg/mL and 80.0μg/mL,relative to the control group were 0.93±0.03 and 0.83±0.08.qRT-PCR showed that the relative expressions ofα-SMA,FN,and ColⅠmRNA in the TGF-β110 ng/mL group were significantly higher than those in the control group(1.62±0.04 vs.0.94±0.05,4.33±0.28 vs.0.98±0.06,2.25±0.97 vs.1.04±0.04,all P<0.05),while the expressions ofα-SMA,FN,and ColⅠmRNA in the Doxy 10.0μg/mL group and Doxy 20.0μg/mL group were significantly lower than those in the TGF-β110 ng/mL group(0.70±0.15,0.41±0.11 vs.1.62±0.04;1.19±0.85,0.79±0.06 vs.4.33±0.28,0.94±0.12,0.52±0.07 vs.2.25±0.97,all P<0.05).Western blotting and immunofluorescence also found the same trend.Conclusion Doxy can exert anti-fibrosis effect by inhibiting the conversion of lung fibroblast induced by TGF-β1.