OBJECTIVE To explore the mechanism of action of Jiedu Fuzheng formula(JDFZ) in anti-hepatocellular carcinoma, based on the method combining network pharmacology and animal experiments.

METHODS Potential therapeutic targets of JDFZ were retrieved from the TCMSP and HERB databases, hepatocellular carcinoma-related targets were collected from GeneCards, OMIM, and DrugBank. The intersection targets of the two were screened to build a protein interaction network(PPI), the intersection targets were imported into the STRING database to remove free protein spots, and the target information of “PPI score>0.9” was imported into Cytoscape 3.10.1 software. The Network Analyzer function was used to analyze the topology structure of the imported data. After the core targets were analyzed and screened, Gene Ontology enrichment(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis were carried out, and a “Drug-Ingredient-Target-Disease” network diagram was constructed. After the relevant data were exported, the mapping analysis was carried out by using the visualization platform. Furthermore, an H22 tumor-bearing mouse model was constructed for experimental validation.

RESULTS Potential therapeutic targets of JDFZ were retrieved from the TCMSP and HERB databases, yielding 342 unique targets. Hepatocellular carcinoma-related targets(18 707) were collected from GeneCards, OMIM, and DrugBank. Intersection analysis identified 314 overlapping targets, and PPI network analysis revealed 22 core targets, including TP53, JUN, SRC, AKT1, MAPK3, and EGFR. Quercetin, luteolin and β-sitosterol were the core components of JDFZ. GO analysis identified 333 significant terms, encompassing 264 biological processes, 25 cellular components, and 44 molecular functions. KEGG pathway analysis highlighted 163 pathways, primarily involving the MAPK signaling pathway, apoptosis pathway, TNF signaling pathway, etc. JDFZ could significantly inhibit tumor growth, significantly reduce tumor weight(P<0.05), and pathology showed significant structural damage to tumor cells. Additionally, JDFZ markedly decreased serum AST and ALT levels and the AST/ALT ratio(P<0.01 or P<0.05). Western blotting analysis revealed significant downregulation of p-EGFR/EGFR and p-ERK/ERK protein expression in tumor tissues(P<0.01).

CONCLUSION This study is based on network pharmacology and H22 tumor-bearing mouse animal experiments. The preliminary results suggest that JDFZ may inhibit liver cancer cell proliferation and reduce liver damage by regulating the EGFR/ERK signaling pathway axis, providing pharmacological evidence for JDFZ treatment of liver cancer.