This study treated HepG2.2.15 cells with Callicarpae Nudiflorae Folium (CNF) water extract (WCNF) and ethanol extract (ECNF), respectively, to evaluate the inhibitory effect of CNF extract on HBV in vitro, and explore the potential mechanism of CNF anti-HBV combined with network pharmacology analysis. CCK-8 method was used in vitro to determine the cytotoxicity of the CNF extract. Hepatitis B e antigen (HBeAg) and hepatitis B surface antigen (HBsAg) antigen secretion were determined by enzyme-linked immunosorbent assay (ELISA), and HBV DNA and key target gene expression were determined by real-time fluorescence quantitative PCR (RT-PCR) method. In the online pharmacological study, the chemical constituents of CNF were searched through literature and TCMSP database, and HBV related targets were searched through Gene Cards database. String database was used to construct protein interaction network, DAVID database was used for GO function and KEGG pathway enrichment analysis, and Cytoscape software was used to construct component-target-pathway network visualization and topological analysis. AutoDock Vina software was used to perform molecular docking analysis of the anti-HBV key components and core targets of CNF, and the key targets were verified by RT-PCR experiment. The results of in vitro experiments showed that the water and ethanol extract could inhibit the secretion of HBeAg and HBsAg antigen and the replication level of HBV DNA, and the inhibitory effect of the water extract was significantly stronger than that of the ethanol extract. Network pharmacology results showed that ursolic acid, quercetin, luteolin, caffeic acid and other compounds were the key active components of CNF anti-HBV effect, tumor protein 53 (TP53), interleukin-6 (IL-6), tumor necrosis factor (TNF), and heat shock protein 90 alpha family class A member 1 (HSP90AA1) are the core targets, involving phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) and mitogen activated protein kinase (MAPK) and other signaling pathways. Furthermore, the results showed that CNF extract inhibited the expression of key HBV genes IL-6, TNF and HSP90AA1, and increased the mRNA expression level of TP53 gene. The above studies revealed that CNF extrect could exert anti-HBV effects through multi-component, multi-target and multi-pathway. In the future, the pharmacodynamic material basis and mechanism of its action will be further discussed based on this study, which will provide a solid theoretical basis for the clinical application and development of new drugs against HBV.