Integrated multi-omics, network pharmacology, and experimental validation reveal that Yinmeikuijie decoction alleviates ulcerative colitis by inhibiting arachidonic acid metabolism.

ETHNOPHARMACOLOGICAL RELEVANCE: Yinmeikuijie decoction (YMKJD), a traditional Chinese herbal formulation, has demonstrated clinical efficacy in alleviating ulcerative colitis (UC). However, its underlying therapeutic mechanisms remain poorly understood.

AIM OF THE STUDY: We aimed to assess the therapeutic efficacy of YMKJD against UC and to unravel its underlying mechanisms through both in vivo and in vitro investigations.

MATERIALS AND

METHODS

Dextran sulfate sodium (DSS)-induced murine UC model mice, TNF-α-treated Caco-2 cells, LPS-stimulated RAW264.7 macrophages, and colon organoids were utilized to investigate YMKJD's therapeutic effects. Macrophage polarization (M1/M2 subsets) was assessed via flow cytometry and immunofluorescence. Immunofluorescence staining and Western blot were performed to evaluate the intestinal epithelial barrier (IEB). Untargeted metabolomics, transcriptomic sequencing, and network pharmacology were integrated to elucidate key pathways. Candidate targets were further verified using qRT‒PCR and Western blot.

RESULTS

YMKJD administration markedly improved clinical symptoms and mitigated colonic tissue damage in UC mice. YMKJD attenuated IEB disruption in vivo and in vitro. Mechanistically, combined bioinformatics and experimental analyses revealed that YMKJD downregulated the expression of the key enzymes (COX-2, 5-LOX, PTGES) in the arachidonic acid (AA) metabolism, thereby suppressing proinflammatory mediators (Leukotriene B4, Prostaglandin E2, ROS) and inhibiting PI3K/AKT signaling in inflamed epithelial cells. Interestingly, YMKJD also blocked COX-2-driven M1 polarization in macrophages, diminishing ROS levels in cocultured colon organoids. Rescue studies confirmed that AA metabolism activation counteracted YMKJD's protective effects in both animal and cellular UC models.

CONCLUSIONS

Our findings demonstrate YMKJD's therapeutic potential and its mechanism in UC, suggesting its utility as a novel treatment strategy.