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The protective effect of Lactobacillus plantarum YS3 (LP‐YS3) on ulcerative colitis (UC) was assessed using a mouse model of dextran sodium sulfate (DSS)‐induced colitis. Different concentrations of LP‐YS4 were administered to the experimental mice by daily gavage. Several inflammatory and biochemical indices, such as interleukin‐2 (IL‐2), interleukin‐10 (IL‐10), interleukin‐6 (IL‐6), interleukin‐1 beta (IL‐1β), tumor necrosis factor‐alpha (TNF‐α), glutathione (GSH), malondialdehyde (MDA), myeloperoxidase (MPO), and nitric oxide (NO), were examined in mouse serum and colon tissue. The mRNA and protein expression levels of c‐Kit, CXC chemokine receptor type 2 (CXCR2), interleukin‐8 (IL‐8), and stem cell factor (SCF) in mouse colon tissue were assessed using Western blot and quantitative polymerase chain reaction (qPCR) assays. The findings indicated that LP‐YS3 remarkably decreased the disease activity index (DAI) of UC mice (p <.05), inhibited colon length shortening induced by UC, and elevated the value of colon weight/length ratio. LP‐YS3 could also markedly reduce (p <.05) the activities of MDA, MPO, and NO; while an increase in the GSH content in the colonic tissue of UC mice. Moreover, LP‐YS3 remarkably increased (p <.05) the serum level of IL‐2 in UC mice, while reduced those of IL‐10, IL‐6, IL‐1β, TNF‐α cytokines. qPCR data revealed that LP‐YS3 could markedly upregulate the expression levels of c‐Kit and SCF, while downregulate those of CXCR2 and IL‐8 in the colonic tissue of UC mice (p <.05). LP‐YS3 exerted an outstanding protective effect on DSS‐induced colitis in C57BL/6J mice, especially at higher concentrations. Practical applications: Lactobacillus plantarum YS3 is a newly isolated and identified lactic acid bacteria. This study confirmed that L. plantarum YS3 can inhibit colitis and has good probiotic potential, which needs further development and utilization. [ABSTRACT FROM AUTHOR]