A Combined Colon Organoid‐Sensory Neuron Model Reveals Epithelial Contribution to Moringin Efficacy Against Painful Inflammatory Bowel Disease

ABSTRACT Visceral pain is a major symptom of inflammatory bowel diseases (IBDs), requiring effective treatment strategies. Gut epithelium, beyond maintaining barrier integrity and microbiota homeostasis, modulates neurosensorial circuitries, influencing visceral sensitivity. Moringa oleifera constituents show beneficial properties that may counteract inflammation‐induced epithelial dysfunction and visceral hypersensitivity. Among these, we investigated moringin (MOR), the isothiocyanate derived from the myrosinase (MYR)‐mediated hydrolysis of glucomoringin (GMG). A three‐stage experimental strategy was applied. First, murine colon organoids were exposed to a pro‐inflammatory cytokine cocktail (CKs; TNF‐α, IL‐1β, IL‐6; 10 ng mL −1 , 6 h) to model epithelial inflammatory stress and treated with MOR (2–30 μM). Second, epithelial–neuronal communication was assessed by exposing primary dorsal root ganglion (DRG) neurons to conditioned media from inflamed organoids, with (CM CKs + MOR 30 μM ) or without (CM CKs ) MOR treatment. Third, the therapeutic relevance of these findings was validated in vivo using a dextran sulfate sodium (DSS)–induced colitis model, in which mice received oral administration of MYR‐bioactivated GMG (GMG + MYR) at different doses (30–100 mg kg −1 ), followed by behavioral and histological assessments. CKs‐treated organoids showed cytotoxicity, increased superoxide dismutase activity, and upregulation of Ccl2 , Ccnd1 , Mki67, and Pyy . MOR 30 μM co‐treatment normalized oxidative stress and gene expression, although cytotoxicity remained unaffected. CM CKs increased c‐Fos and CGRP expression in DRG neurons, while CM CKs + MOR 30 μM prevented this response, indicating MOR modulation of epithelial inflammation‐driven neuronal activation. In DSS‐treated mice, oral GMG (30–100 mg kg −1 ) + MYR dose‐dependently reduced visceral hypersensitivity, pain‐associated behavioral alterations, and colonic damage. Overall, MOR/GMG + MYR showed gut‐protective and analgesic effects under intestinal inflammation. Intestinal organoids effectively model inflammation and therapeutic responses, potentially reducing animal use in IBD research.