The development of inflammatory bowel disease (IBD) is closely linked to inflammatory damage and dysbiosis. Recently, probiotics are being increasingly used to improve intestinal health. Probiotic-based therapies can prevent IBD by restoring the balance of gastrointestinal microbiota, reducing gut inflammation, and increasing the concentration of short-chain fatty acids (SCFAs). The present study aimed to investigate the protective effects of Lactiplantibacillus plantarum subsp. plantarum Dad-13, a novel probiotic strain derived from dadih (Indonesian curd from buffalo milk), on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in BALB/c mice. The results showed that probiotic Dad-13 supplementation at a dose of 10(7) or 10(9) CFU/mL improved the clinical symptoms of IBD and enhanced the production of SCFAs, particularly propionate and butyrate. Moreover, probiotic Dad-13 supplementation significantly decreased the levels of pro-inflammatory cytokines [tumor necrosis factor-alpha, interleukin (IL)-6, and IL-1beta] and significantly increased the levels of anti-inflammatory cytokines (IL-10). These findings show that L. plantarum Dad-13 can effectively prevent TNBS-induced colitis by modulating SCFA production and inflammatory cytokines.