EGCG抑制HBV复制的分子机理研究

Molecular mechanism of epigallocatechin gallate-mediated anti-HBV replication effect

目的研究表没食子儿茶素没食子酸酯（epigallocatechin gallate, EGCG）抑制乙型肝炎病毒（hepatitis B virus, HBV）复制的分子机理。方法利用CCK-8测定细胞的活力。利用荧光定量PCR方法检测细胞内外HBV DNA的含量。利用Western Blot检测HNF4a的表达及siRNA对HNF4a的敲除作用。结果本研究发现，利用EGCG处理HepG2.2.15细胞可显著降低细胞内外HBV DNA的含量，但不影响细胞的活性，表明EGCG具有抑制HBV复制的作用。前人研究发现，肝细胞因子4a（HNF4a）是一个促进HBV复制的重要细胞因子。本研究中发现，EGCG可显著下调HepG2.2.15细胞中HNF4a的表达。沉默HNF4a表达后，EGCG抑制HBV DNA复制的作用显著下降。结论上述结果提示，在HepG2.2.15细胞中，通过下调HNF4a的表达，从而抑制HBV DNA复制，可能是EGCG发挥其抗HBV作用的一条重要途径。

ObjectiveTo elucidate the mechanism of epigallocatechin gallate （EGCG） mediated anti-HBV effect. MethodsThe CCK-8 kit was used to test cell viability in response to EGCG treatment. For HBV DNA replication assay, purified HBV DNA was analyzed by real-time PCR assay. Western blotting was used to confirm HNF4α expression in response to EGCG or siRNA treatment.ResultsOur result showed that, EGCG treatment significantly decreasee HBV DNA level both in vivo and in vitro without affecting cell viability. Curiously, we found that EGCG treatment downregulated HNF4α expression. Considering that HNF4α could restrain HBV replication, we knocked down HNF4α in HepG2.2.15 cells and found that the response of HBV replication to EGCG decreased obviously.ConclusionsThe above result suggest that HNF4α could be an important intracellular factor in EGCG mediated anti-HBV signaling pathway.