Inflammatory bowel disease (IBD) involves chronic gut inflammation and barrier damage. While natural flavonoids show promise for IBD treatment, cirsiliol's effects were unknown. This study examined cirsiliol's therapeutic potential in mice with DNBS-induced colitis and its mechanisms in vitro and in vivo. Mice received either ethanol (vehicle), DNBS, or cirsiliol (10 or 30 mg/kg). Disease severity was measured by weight loss, colon shortening, and Disease Activity Index (DAI). Lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma) stimulated rat intestinal cells (IEC-6) modeled inflammation in vitro. Oxidative stress, inflammatory factors, barrier proteins (Claudin-1, Occludin, E-cadherin), and NF-kappaB/MAPK pathways were analyzed. Cirsiliol significantly alleviated colitis symptoms, reducing weight loss and DAI while improving colon length. It suppressed pro-inflammatory cytokines (TNF-alpha, IL-6, IL-1beta), boosted antioxidants (increased GSH, decreased MDA), and restored tight junction proteins. Cirsiliol also reduced apoptosis and enhanced cell migration. Mechanistically, it inhibited the activation of NF-kappaB and MAPK pathways seen in DNBS-induced mice. These findings demonstrate cirsiliol's protective effects against colitis via anti-inflammatory, antioxidant, and barrier-enhancing actions, mediated by NF-kappaB and MAPK pathway inhibition, suggesting its potential as a novel IBD treatment.