Unveiling the Therapeutic Mechanism of Epimedium Herba on Prostate Cancer through Network Pharmacology and Experimental Validation

Objective: Epimedium Herba (EH) has significant therapeutic potential in alleviating prostate cancer (PC), but its mechanisms of action remain unclear. This study aims to evaluate the pharmacological mechanisms of EH in treating PC through network pharmacology analysis and experimental validation. Methods: Active components and potential targets of EH were screened using network pharmacology from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). The STRING database was used to construct a protein-protein interaction (PPI) network for shared targets and identify core anti-PC targets. mRNA and protein expression of core target genes in normal prostate and PC tissues, as well as their relationship with overall PC survival, were assessed using The Cancer Genome Atlas (TCGA), Human Protein Atlas (HPA), and Gene Expression Profiling Interactive Analysis (GEPIA) databases. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted on the potential targets. Molecular docking of quercetin with key targets (TP53, TNF, HSP90AA1, AKT1, CASP3, ESR1) was performed, and results were visualized using PyMOL. Finally, in vitro experiments were conducted to validate the predictions made by network pharmacology. Results: Twenty-three active ingredients of EH were identified, and the intersection of potential targets with PC targets yielded 183 potential targets. PPI network analysis identified six key genes, including TP53, TNF, HSP90AA1, AKT1, CASP3, and ESR1. GO enrichment analysis revealed 2369 biological processes, 77 cellular components, and 215 molecular functions. KEGG pathway enrichment analysis showed that the anti-cancer effects of EH were mediated by the IL-17, TNF, PI3K-Akt, apoptosis, p53, HIF-1, MAPK, NF-κB, and EGFR tyrosine kinase inhibitor resistance pathways. Validation of the core targets indicated that the experimental results were consistent with the study’s findings. Molecular docking showed stable binding of the six core targets with quercetin. In vitro experiments demonstrated that quercetin inhibited proliferation and induced apoptosis in ACT-1 cells. Conclusion: This study identifies potential therapeutic targets for PC through network pharmacology and experimental validation.