ScopeAt present, the incidence rate of ulcerative colitis (UC) continues to increase, causing a global burden. In addition, therapeutic drugs have great side effects. According to modern pharmacology, agarwood leaves have anti‐inflammatory, antibacterial, hypoglycemic, and lipid‐lowering effects. Therefore, this experiment on DSS induced colitis treatment of polyphenolic substances in agarwood leaves is feasible and in line with the current hot topic of using natural substances instead of drugs for treatment.Methods and resultsALP supplementation promotes the expression of tight junction proteins occludin and Zonula occludens protein 1 (ZO‐1) on colonic tissues, repairs the intestinal barrier, and relieves further colonic tissue damage. Besides, ALP effectively inhibits the activation of nuclear factor kappa‐B (NF‐кB) signaling pathway and reduces the release of proinflammatory cytokines. Moreover, ALP reverses the alteration of gut microbiota in the colitic mice by increasing the abundances of Parabacteroides, Chlamydia, and Lachnospiraceae, and decreasing the abundances of Bacteroides and Phocaeicola. Furthermore, the correlation analysis suggested that ALP can attenuate DSS‐induced UC, which is probably related to the alterations in the gut microbiota.ConclusionALP can ameliorate DSS‐induced UC by modulating gut microbiota, intestinal barrier function, and inflammatory responses.