ppGalNAc-T2基因下调对肝星状细胞TGFβ1-Smad3激活途径基因表达的影响

The effect of knock-down of ppGalNAc-T2 on genes expression in TGFβ1-Smad3 activation pathway in HSC

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 observe the influence of ppGalNAc-T2 on genes expression in TGFβ1-Smad3 activation pathway in rat hepatic stellate cells（HSC）.Methods After the transfection of plasmid of siRNA in HSC,the mRNA expression of genes related to TGFβ1-Smad3 activation pathway was detected by RT-PCR and the Smad3 protein was detected by Western blot and immunofluorescence.Results Compared with that in the normal cells,the knock-down of ppGalNAc-T2 had no significant effect on the mRNA expression of Smad 2,Smad 3,Smad 4,Col3α1,Tbrg4,TGFβ,TGFβ2 and TGFβr3 and the protein expression of Smad3（P0.05）.However,the mRNA expressions of TGFβr1 and TGFβr2 were significantly down-regulated（P0.05）.Conclusion Many gene expressions in TGFβ1-Smad3 activation pathway in HSC are not varied after the ppGalNAc-T2 is down-regulated,whose function in HSC should be studied further.