Integrated Multi-Omics, Machine Learning, and Single-Cell Analysis Provide Insights into Potential Multi-Target Candidate Mechanisms of Combined MAGL11 and Icaritin in Alleviating Imidazole Propionate-Driven Metabolic Intestinal Injury.

Background: Unhealthy diets, especially high red meat intake, induce gut dysbiosis and abnormal accumulation of a harmful histidine metabolite imidazole propionate (IMP), thereby promoting ulcerative colitis (UC) development. Although combined treatment with a MAGL inhibitor (MAGL11) and icaritin (Y003) exerts combined therapeutic effects in the classical DSS‑induced UC model, its protective effects and molecular mechanisms against IMP‑driven metabolic model remain unclear. Purpose: The present study was designed to evaluate the therapeutic effect of MAGL11 and Y003 on IMP‑driven metabolic intestinal injury model and to elucidate the associated molecular mechanisms. Methods: We evaluated the protective effects of MAGL11 and Y003 combination therapy in an IMP-induced intestinal injury model and integrated multi‑omics data to explore the underlying mechanisms. Results: In this short-term preclinical study in male mice, the combined treatment significantly alleviated UC symptoms, suppressed pro‑inflammatory cytokines, repaired the intestinal barrier, corrected metabolic disturbances, and reshaped the gut microbiota. Four hub genes were prioritized by machine learning integrating differential expression analysis and WGCNA of GEO data together with targets of MAGL11+Y003 and fecal metabolites. Molecular docking predicted favorable binding affinities between MAGL11/Y003 and the four target proteins (CA2, ABCB1, ABCG2, MMP3). Single‑cell analysis revealed cell‑type‑specific dysregulation of these genes. Furthermore, CA2, ABCB1, and ABCG2 were downregulated, while MMP3 was upregulated in the colon tissues of IMP‑induced model mice, while the combination therapy reversed these changes. Proteomics and Western blot validation indicated that the therapeutic efficacy of the combination treatment was associated with the suppression of the VEGF and MAPK pathway as well as the modulation of apoptosis- and metabolism-related pathways. Conclusion: Collectively, MAGL11 and Y003 combined therapy alleviates IMP‑driven metabolic intestinal injury through multi‑target, multi‑pathway actions with a greater therapeutic effect, offering a novel strategy for metabolically associated colitis and prioritizing CA2, ABCB1, ABCG2, and MMP3 as candidate treatment-responsive molecules requiring further functional validation.