Synergistic Suppression of Inflammation by AZA, Rice Bran Oil, and AntimiR-466l through IL-6/NF-κβ/TLR4 Pathways in Mice Model of Ulcerative Colitis

This study aimed to evaluate the anti-inflammatory effects of azathioprine (AZA), rice bran oil (RBO), and antisense miR-466l in a dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) model. The synergistic effect of AZA with RBO and the therapeutic potential of miR-466l inhibition were investigated. Forty male C57BL/6 mice were assigned to six groups: control, UC, UC + AZA, UC + RBO, UC + AZA + RBO, and UC + anti-miR-466l. Disease severity was evaluated by colon length, Disease Activity Index (DAI), and histopathological scoring. Expression levels of IL-6/IL-10/NF-κβ/TLR4 and miR-466l were analyzed using qRT-PCR. DSS exposure resulted in severe inflammation, colon shortening, and increased DAI scores, along with upregulation of IL-6/TLR4/NF-κβ, and miR-466l, and a marked reduction in IL-10. AZA and RBO treatments attenuated these effects, improving mucosal architecture and restoring cytokine balance. The combined AZA + RBO therapy exhibited the strongest anti-inflammatory response, significantly suppressing pro-inflammatory markers and enhancing IL-10 expression. Anti-miR-466l treatment improved both molecular and histological parameters, highlighting its role in inflammation regulation. The combination therapy provided superior outcomes compared to monotherapies, suggesting a synergistic modulation of immune and oxidative pathways. Overall, integrating AZA, RBO, and miR-466l inhibition may offer a promising and synergistic therapeutic strategy for ulcerative colitis management.