Functional molecules from Ganoderma resinaceum fermentation broth for cosmetics: LC–MS/MS and in silico–to–in vitro evidence for anti-inflammatory activity and skin repair–relevant effects

Background Ganoderma resinaceum fermentation broth is a complex natural mixture with potential skin-soothing and repair-related effects, but its active constituents and mechanisms have not been systematically clarified. Methods This study investigated the skincare-related bioactivities and mechanisms of G. resinaceum fermentation broth using an integrated strategy combining LC-MS/MS metabolomic profiling, network pharmacology, molecular docking, molecular dynamics simulations, and cell-based assays. Compound targets predicted by Super-PRED were intersected with skin-related genes from GeneCards and UniProt, followed by protein-protein interaction analysis, GO/KEGG enrichment, and compound-target-pathway network construction. Anti-inflammatory and repair-related effects were evaluated in LPS-stimulated Ana-1 macrophages and HaCaT keratinocytes. Results LC-MS/MS tentatively annotated 81 small molecules, mainly including amino acids, sugars, and triterpenes. Network analysis identified 141 putative skin-related targets and 19 relevant pathways, notably PI3K-Akt, MAPK, and TNF signaling. Docking against TNF-α (PDB ID: 2E7A) showed favorable binding for ten prioritized metabolites, with affinities ≤ −5 kcal/mol; ganoderic acids A, B, C2, and glycoursodeoxycholic acid showed the strongest affinities (−10.8 to −9.3 kcal/mol). Molecular dynamics simulations supported relatively stable protein-ligand interactions under the present simulation conditions. In LPS-stimulated Ana-1 macrophages, individual compounds at 1 mg/L reduced TNF-α levels by 16.77%-18.29%, whereas 0.1% (v/v) fermentation broth reduced TNF-α by 28.11%. In HaCaT keratinocytes, scratch closure was enhanced by 1.67-5.68-fold within 24 h under non-cytotoxic conditions. Notably, the fermentation broth showed stronger TNF-α suppression than the tested representative compounds, suggesting combined contributions from multiple constituents and possibly additional untested trace-active components. Conclusions G. resinaceum fermentation broth exerts multi-component, multi-target, and multi-pathway actions that attenuate inflammation and promote repair. Its stronger anti-inflammatory activity relative to the tested representative compounds suggests that the biological effect of the whole fermentation system may arise from integrated contributions of multiple constituents, providing a scientific basis for its development as a skincare ingredient.