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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。
Objective To investigate the effect of hepatitis B virus e antigen (HBeAg) on Toll-like receptor 3 (TLR3) expression in Bewo cells. Methods Bewo cells were treated with TLR3 ligand polyI: C first and the dynamic changes of TLR3 mRNA expression were observed. Then, 2μg and 8μg HBeAg recombinant plasmid pcDNA3.1 (+) - HBe were transfected into Bewo cells. After 48 h, the cells were treated with TLR3 ligand polyI: C for 12 h. Finally, Bewo cells were treated with different concentrations of IFN-β for 12 h. Real-time fluorescent quantitative RT-PCR and ELISA were used to detect the expression of TLR3 mRNA and the level of IFN-β in the supernatant. Results PolyI: C significantly induced the expression of TLR3 mRNA (P <0.05 or 0.001) in Bewo cells in a time and dose dependent manner. Compared with the control group, the polyI: C transfected 8μg HBeAg recombinant plasmid induced the expression of TLR3 in Bewo cells and The level of IFN-β was significantly decreased (P <0.001). IFN-β significantly induced the expression of TLR3 mRNA (P <0.001) in Bewo cells in a dose-dependent manner. Conclusion HBeAg may down-regulate the expression of TLR3 mRNA in Bewo cells by down-regulating the production of IFN-β, providing a new approach for the prevention and treatment of HBV intrauterine infection.