摘要
目的观察wnt抑制因子-1(WIF)对转化生长因子β1(TGF-β1)活化肝星状细胞(HSC)的影响。方法构建真核表达质粒pEGFP-C1-WIF并通过脂质体介导方法,将pEGFP-C1-WIF转染体外培养的HSC-T6细胞株,用TGF-β1刺激转染(试验组)和未转染的HSC-T6细胞株(对照组),并用逆转录PCR(RT-PCR)和Western blot法检测各组细胞(包括正常细胞组)smad3、β-连环蛋白(β-catenin)mRNA及其蛋白质的表达。结果试验组HSC-T6细胞株smad3、β-catenin mRNA及蛋白质表达明显低于正常组HSC-T6细胞株,更低于对照组HSC-T6细胞株(P<0.05)。β-catenin和smad3蛋白质的表达均与α-肌动蛋白(α-SMA)蛋白质的表达显著相关(r=0.800,P<0.01;r=0.743,P<0.01)。结论 WIF能通过下调wnt/β-catenin以及TGF-β1/smads信号通路的生物学效应抑制HSC-T6细胞的活化,从而延缓肝纤维化的发展。
Objective To observe the effect of Wnt inhibitory factor（WIF） on the activation of hepatic stellate cells stimulated by transforming growth factor-beta1. Methods The recombinant expression plasmid pEGFP-C1-WIF was constructed and transfected into cultured HSC-T6 cells. HSC-T6 cells were incubated with TGF-β1 before and after transfection. The expression of smad3 and β-catenin mRNA and smad3,α-SMA,β-catenin protein in HSC-T6 cells （including the normal group） were evaluated by PT-PCR and Western-blot respectively. Results The expressions of smad3 and β-catenin mRNA and protein in HSC-T6 cells with WIF were both lower than those in the normal control cells. The expression of β-catenin and smad3 protein had a positive correlation with the level of α-SMA protein in cells（r=0.800,P〈0.01; r=0.743,P〈0.01,respectively）. Conclusion WIF could suppress the activation process of HSC-T6 and might delay the development of hepatic fibrosis through inhibiting the wnt/β-catenin and TGF-β1/smads pathways.
出处
《肝脏》
2010年第5期338-341,共4页
Chinese Hepatology
基金
安徽省教育厅基金(KJ2010B404)
