Abstract

Ethnopharmacological relevance Herba Euphorbiae Humifusae ( Euphorbia humifusa Willd., EH), named Dijincao , a traditional Chinese herbal medicine used to treat dysentery and enteritis, shows therapeutic potential for ulcerative colitis (UC), yet its mechanism remains unclear. Aim of the study To elucidate the efficacy of EH against UC and its mechanism. Materials and methods The effect of EH extract (EHE) on UC was evaluated using DSS-induced mice. Integrated analysis of metabolomics and network pharmacology based on the colonic exposure of prototypes in EHE predicted key targets and pathways involved in the treatment of UC, while molecular docking and biological experiments investigated the mechanism. Spearman’s correlation analysis identified key anti-UC active components, followed by in vitro activity and mechanistic validation in RSL3-induced HCT116 cells and intestinal organoid models. Results EHE alleviated UC mice in a dose-dependent manner. Network pharmacology predicted the major targets of the 18 colonic exposure components in EHE to be signaling pathways involved in inflammation and lipid metabolism. Metabolomics revealed that EHE primarily regulated glutathione metabolism and arachidonic acid metabolism. Through the integrated analysis of network pharmacology and metabolomics, suggesting that lipid peroxidation involving 11 pivotal targets may mediate the beneficial effect of EHE against UC, while molecular docking analysis and experiments substantiated that the amelioratory action of EHE partially contributed to its inhibition on ACSL4/COX-2 axis mediated lipid peroxidation. Notably, quercetin, kaempferol and naringenin, as key active constituents of EHE against UC, demonstrated the direct regulatory effect on ACSL4/COX-2 axis mediated lipid peroxidation in vitro , which was attenuated upon treatment with overexpressed ACSL4. Conclusions This study demonstrated that EH ameliorates UC by modulating ACSL4/COX-2 axis mediated lipid peroxidation, the