乙型肝炎病毒e抗原对Bewo细胞Toll样受体3表达的影响

目的探讨乙型肝炎病毒e抗原（HBeAg）对Bewo细胞Toll样受体3（TLR3）表达的影响。方法首先用TLR3配体polyI∶C处理Bewo细胞,观察细胞TLR3 mRNA表达的动力学变化。然后将2μg和8μg HBeAg重组质粒pcDNA3.1（＋）-HBe转染Bewo细胞,48 h后,用TLR3配体polyI∶C处理12 h。最后,用不同浓度的IFN-β处理Bewo细胞12 h。采用实时荧光定量RT-PCR和ELISA分别检测细胞TLR3 mRNA表达及细胞上清IFN-β水平。结果 polyI∶C可显著诱导Bewo细胞TLR3 mRNA表达（P〈0.05或0.001）,且呈时间和剂量依赖性;与对照组相比,转染8μg HBeAg重组质粒组polyI∶C诱导Bewo细胞TLR3表达及IFN-β水平显著下降（P〈0.001）,IFN-β可显著诱导Bewo细胞表达TLR3 mRNA（P〈0.001）,且呈剂量依赖性。结论 HBeAg可能通过下调IFN-β的产生而抑制Bewo细胞TLR3 mRNA的表达,为防治HBV宫内感染提供了新的途径。

Effects of HBeAg on the Expression of Toll-like Receptor 3 in Bewo Cells

Objective To study the effect of HBeAg on the expression of Toll-like receptor 3（TLR3） in Bewo cells..Methods Firstly,Bewo cells were exposed to TLR3 ligand polyI ∶C to observe the kinetic change of TLR3 mRNA expression in Bewo cells.Secondly,2 μg and 8 μg HBeAg recombinant vector pcDNA3.1（＋）-HBe was transfected into Bewo cells.48 h after transfection,the cells were treated with polyI ∶C for 12 h.Eventually,Bewo cells were stimulated with different concentrations of IFN-β for 12 h.TLR3 mRNA expression and IFN-β level in the culture supernatant were determined by real-time fluorescence quantitative RT-PCR（qRT-PCR） and ELISA,respectively.Results polyI∶C and IFN-β could significantly induce TLR3 mRNA expression in Bewo cells（P0.05 or 0.001）in a time-and dose-dependent manner.Compared with the control,the levels of polyI∶C-induced TLR3 mRNA and IFN-β production in cells transfected with 8 μg HBeAg recombinant vector group were strikingly decreased（P0.001）.IFN-β could significantly induce TLR3 mRNA expression in Bewo cells（P0.001）in a dose-dependent manner.Conclusion HBeAg may inhibit the expression of TLR3 mRNA through down-regulating IFN-β in Bewo cells.Results from this study will provide a new method for prevention and cure intrauterine HBV infection.