To explore the potential anti-melanoma mechanisms of juglone using network pharmacology combined with experimental validation.Methods:Potential targets of juglone and melanoma were screened using databases including the Comparative Toxicogenomics Database(CTD)，Therapeutic Target Database(TTD)，Online Mendelian Inheritance in Man(OMIM)，GeneCards，and DisGeNET.The intersecting targets were analyzed using the STRING database，and a protein-protein interaction(PPI) network was constructed via Cytoscape.Gene Ontology(GO) enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses were performed using the Metascape platform.Cellular experiments were conducted to assess the effects of juglone on B16F10 melanoma cell viability，invasion ability，and the levels of inflammatory cytokines interleukin-1β(IL-1β)，interleukin-6(IL-6)，and tumor necrosis factor-α(TNF-α) in the cell supernatant，as well as the expression of tumor protein p53(p53).Results:A total of 37 intersecting targets were identified.The top three core targets were tumor protein p53，cysteine aspartic protease 3(CASP3)，and TNF.KEGG pathway enrichment analysis revealed that these targets are closely related to cancer-related signaling pathways.GO enrichment analysis showed that biological processes were mainly involved in responses to xenobiotic stimuli.Cell experiments demonstrated that juglone inhibited B16F10 cell proliferation and invasion，suppressed the expression of inflammatory cytokines，and upregulated p53 and phosphorylated p53(p-p53) protein expression.Conclusion:Juglone exerts anti-melanoma effects through multiple targets and pathways，providing scientific evidence and experimental support for further investigation of its mechanisms in inhibiting tumor invasion and metastasis.