This study investigates the role of humic acid (HA) in attenuating Dextran sulfate sodium (DSS)-induced UC in male Wistar rats. Twenty male Wistar rats were randomly assigned to groups (n = 5). Group 1 served as controls; Group 2 received 5% DSS alone; Group 3 received 5% DSS followed by humic acid (30 mg/kg); and Group 4 received 5% DSS followed by sulfasalazine (200 mg/kg). DSS was administered orally to induce colitis in Groups 2, 3, and 4. Colitis was induced for five days and drug treatment done for another 5 days Disease activity index was assessed on days 1, 3, 5, and 10. Animals were euthanized by cervical dislocation, colon specimens harvested for macroscopic assessment, histological and biochemical assays. Data were analyzed using one way ANOVA (p < 0.05). Results show humic acid treatment significantly attenuated DSS-induced colitis by reducing inflammation markers (TNF-α, interleukin-6, interleukin-1β, and arginase), lipid peroxidation (MPO) and restoring colonic histological integrity. Significant improvements were observed in colonic tissue nitrite levels and Muc-2 expression. Conclusively, HA shows promise as an alternative therapeutic for UC, offering antioxidant, anti-inflammatory, and mucosal barrier-protective effects.