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Standardized ethanolic extract of the rhizome of Curcuma xanthorrhiza (CE) with xanthorrhizol was evaluated for anti-inflammatory activity in a mouse model of 5% dextran sulfate sodium (DSS)-induced ulcerative colitis (UC). Treatment with CE (4 or 40 mg/kg) attenuated the disease activity index, shortening of the colon, histological changes, and myeloperoxidase activity. These changes were associated with marked inhibition of inflammation-related expression of genes for proinflammatory cytokines, matrix metalloproteinase and its inhibitor, and acute-phase proteins. Notably, CE also inhibited DSS-activated release of nuclear factor-κB (NF-κB) from its inhibitor proteins, called inhibitors of NF-κB (IκB), and reduced the level of expression of cyclooxygenase-2 and serum S100 calcium-binding protein a8. Our results show that CE has an ameliorating effect against DSS-induced UC possibly through its anti-inflammatory action in the colon, and may be an attractive natural source for the development of functional food to modulate colitis.