Objective: To elucidate the mechanism of epigallocatechin gallate (EGCG) mediated anti-HBV effect. Methods: The 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. Results: Our 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. Conclusions The above result suggest that HNF4α could be an important intracellular factor in EGCG mediated anti-HBV signaling pathway.