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目的探讨糖基转移酶ppGalNAc-T2对大鼠肝星状细胞的转化生长因子(TGF)β1-Smad3激活的影响。方法在大鼠肝星状细胞中转染ppGalNAc-T2的siRNA表达质粒,用RT-PCR检测相关基因mRNA表达,Western blot以及免疫荧光方法检测Smad3基因蛋白表达。结果相对于正常细胞,ppGalNAc-T2下调后,肝星状细胞中Smad2、Smad3、Smad4、胶原3α1(Col3α1)、转化生长因子β调节基因4(Tbrg4)、TGFβ、TGFβ2及转化生长因子β受体(TGFβr)3的mRNA及Smad3的蛋白表达几乎没有变化(P>0.05),但TGFβr1和TGFβr2都有较大程度的降低(P<0.05)。结论在大鼠肝星状细胞中,ppGalNAc-T2下调对于TGFβ1-Smad3激活途径基因表达基本没有影响,其在肝星状细胞中的具体功能有待于进一步研究。
Objective To investigate the effect of glycosyltransferase ppGalNAc-T2 on the activation of transforming growth factor β1-Smad3 in rat hepatic stellate cells. Methods The siRNA expression plasmids of ppGalNAc-T2 were transfected into rat hepatic stellate cells. The expression of mRNA was detected by RT-PCR. The protein expression of Smad3 was detected by Western blot and immunofluorescence staining. Results Compared with normal cells, the expression of Smad2, Smad3, Smad4, Collagen 3α1, Tbrg4, TGFβ, TGFβ2 and TGFβ in hepatic stellate cells were decreased after ppGalNAc- (P <0.05). However, TGFβr1 mRNA and Smad3 protein expression were almost unchanged (P> 0.05), but both TGFβr1 and TGFβr2 were significantly decreased (P <0.05). Conclusion In rat hepatic stellate cells, the downregulation of ppGalNAc-T2 has no effect on the gene expression of TGFβ1-Smad3 activation, and its function in hepatic stellate cells needs further study.