ETHNOPHARMACOLOGICAL RELEVANCE: Xipayi Kuijie'an herbal enema preparation, a traditional Uyghur medicinal preparation derived from Quercus infectoria galls (QIG), has been clinically employed at Xinjiang Uyghur Medical Hospital for ulcerative colitis (UC) management. Despite its well-documented therapeutic efficacy, the precise mechanisms underlying its pharmacological actions remain poorly understood. AIM OF STUDY: This study aimed to elucidate the therapeutic mechanisms of QIG against UC through an integrated approach combining network pharmacology analysis with experimental validation. MATERIALS AND METHODS: An integrated approach combining network pharmacology analysis and molecular docking simulations was employed to identify bioactive compounds and their corresponding molecular targets. The effects of QIG on dextran sodium sulfate (DSS) induced colitis were systematically investigated, including disease activity index (DAI), colon length, histopathological observations, as well as the determination of colonic mucosal barrier permeability and biochemical indicators. RESULTS: QIG significantly improved DAI and CMDI scores, reduced colonic shortening, decreased colonic mucosal barrier permeability, reduce the release of pro-inflammatory factors and mitigated histopathological changes. CONCLUSION: This study demonstrates the efficacy of an integrated approach combining network pharmacology with experimental validation as a robust strategy for elucidating the therapeutic mechanisms of traditional medicines, as exemplified by the systematic investigation of QIG's anti-ulcerative colitis properties.