丹蒽醌通过MAPK-HNF4α信号通路抑制乙型肝炎病毒复制的作用机制研究

Study on the mechanism of danthron in inhibiting hepatitis B virus replication through the MAPK-HNF4α signaling pathway

目的:本课题旨在研究丹蒽醌对HBV复制的抑制作用及其在体外抗HBV的分子机理。方法:运用MTT法检测丹蒽醌对Hep G2.2.15细胞的细胞毒性;利用Q-PCR检测丹蒽醌对Hep G2.2.15细胞内HBV DNA表达量的抑制效果;利用蛋白免疫印迹实验检测丹蒽醌对Hep G2.2.15细胞内蛋白信号的调控作用。结果:丹蒽醌在不影响Hep G2.2.15细胞增殖的浓度下就能够显著抑制其HBV cccDNA的复制,IC_(50)约为20.43μM。分子层面研究表明,丹蒽醌可以通过激活MAPK通路,从而降低乙肝复制相关的HNF4α蛋白的表达,同时还能下调乙肝复制相关的HBx表达从而有效阻止HBV的复制。结论:丹蒽醌通过激活MAPK1/3通路从而抑制Hep G2.2.15细胞中HBV的复制。

Objective:This project aimed to investigate the inhibitory effect of danthron on HBV replication and its molecular mechanism against HBV in vitro.Method:The cytotoxicity of danthron on HepG2.2.15 cells was detected by MTT method.Q-PCR was used to detect the inhibitory effect of danthron on the expression of HBV DNA in HepG2.2.15 cells.The effect of danthron on intracellular HBeAg in HepG2.2.15 cells was detected using ELISA assay.Western blot experiments were used to detect the regulatory effect of danthron on HepG2.2.15 intracellular protein signaling.Result:Danthron significantly inhibited HBV cccDNA replication at concentrations that do not affect the proliferation of HepG2.2.15 cells,with an IC_(50) of approximately 20.43μM.Danthron can activate the MAPK pathway,thereby reducing the expression of HNF4a,while also downregulating HBx expression to effectively prevent HBV replication.Conclusion:Danthron inhibits the replication process of HBV in HepG2.2.15 cells by inhibiting the expression of HBx and HNF4αproteins.