β-连环蛋白对转化生长因子β 1活化HSC-T6细胞的影响

目的 观察β-连环蛋白对转化生长因子β 1(TGF β1)活化肝星状细胞(HSC)的影响. 方法通过脂质体介导,将β-连环蛋白真核表达质粒pcDNA3.1(+)-β-catenin]和绿色荧光质粒pEGFP-N1共转染体外培养的HSC-T6细胞株,用逆转录PCR和Western blot法检测经TGF β1刺激后两组细胞smad3、β-连环蛋白mRNA及蛋白质的表达.组间比较采用方差分析.结果 β-连环蛋白转染阳性的HSC-T6细胞株经TGF β 1刺激后表达smad3、β-连环蛋白及其mRNA明显强于未转染β-连环蛋白基因的HSC-T6细胞,更强于正常培养的HSC-T6细胞,smad3mRNA相对表达量分别为;0.642±0.011、0.501±0.021、0.511±0.019、0.356±0.017,F=135.304,P<0.05;β-连环蛋白mRNA相对表达量分别为:0.783±0.021、0.543±0.033、0.538±0.024、0.212±0.019,F=267.340,P值均<0.05.smad3蛋白质相对表达量分别为:0.892±0.012、0.124±0.011、0.130±0.021、0.003±0.001,F=2823.813,P<0.05;β-连环蛋白相对表达量分别为:0.921±0.020、0.210±0.010、0.208±0.008、0.002±0.001,F=3440.982,P<0.05.β-连环蛋白和smad3蛋白质的表达均与α-平滑肌肌动蛋白的表达显著相关(r=0.901,P<0.01;r=0.939,P<0.01).结论 β-连环蛋白基因转染体外培养的HSC-T6细胞株,促使HSC表达smad3、α-平滑肌肌动蛋白增强,提示β-连环蛋白能增强TGF β1的致纤维化作用.

Effect of beta-catenin on the activation of hepatic stellate cells induced by transforming growth factor-betal

Objective To investigate the effect of beta-catenin on the activation of hepatic fibrosis by transforming growth factor-beta 1 (TGF β1). Methods The recombinant expression plasmids pcDNA3.1 (+)-beta-catenin and pEGFP-N1 were cotransfected into cultured HSC-T6 cells.The expression of smad3, beta-catenin and alpha-SMA, beta-catenin protein in TGF β1 treated HSC-T6 cells were detected by RT-PCR and Western-blot. Results The expression of smad3 and beta-catenin in the co-transfected cells was higher than that in the untransfected cells (smad3 mRNA were 0.642±0.011, 0.501±0.021, 0.511±0.019, 0.356±0.017, respectively, F=135.304, P<0.05. Beta-catenin mRNA were 0.783±0.021, 0.543±0.033, 0.538±0.024, 0.212±0.019, respectively, F=267.340, P<0.05. Smad3 protein were 0.892±0.012, 0.124±0.011, 0.130±0.021, 0.003±0.001, F=2823.813, P<0.05. Beta-catenin protein were 0.921± 0.020, 0.210±0.010, 0.208±0.008, 0.002±0.001, respectively, F= 3440.982, P<0.05). The expression of beta-catenin and smad3 protein had a positive correlation with the level of alpha-SMA protein in cells (r=0.901, P<0.01; r=0.939, P<0.01). Conclusions Expression of smad3/alpha-SMA/beta-catenin is increased in the cultured HSC-T6 cells transfected by beta-catenin gene, especially when the transfected cells are stimu-lated by TGF β1. Our data suggest that beta-catenin could aggravate hepatic fibrosis induced by TGF β1.