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However, its preventive potential against intestinal inflammation remains to be poorly understood. This study explored the protective effects of arabinoxylan from barley, rice bran, and highland barley on colitis mice. The results suggested that all arabinoxylan significantly ameliorated weight loss, disease activity index (DAI), colon length, and inflammatory cytokine levels in colitis mice. Notably, arabinoxylan interventions increased Bacteroides abundance while reducing Clostridium and Escherichia-Shigella abundance, thereby improving the microbiota composition. Metabolomics revealed that arabinoxylan from all three sources modulated colonic metabolism in colitis mice, particularly enriching the alanine, aspartate, and glutamate metabolic pathways. 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