Abstract: Objective:To investigate the mechanism of tributyrin (TB) in alleviating 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced intestinal inflammation in rats with experimental colitis. Methods:A total of 24 male Sprague Dawley rats were randomly divided into four groups (6 rats in each group):control group, model group (TNBS group), low-dose TB group (TNBS+TB 600 mg/kg) and high-dose TB group (TNBS+TB 1,000 mg/kg), and the low-dose and high-dose TB groups were gavaged with the corresponding dose of TB for one week before and after modeling. After modeling, the rats were observed for activity and fecal characteristics, the rats were assessed for the disease activity index (DAI), and the colonic tissues were observed and stained with hematoxylineosin and scored for histological damage after the rats were put to death. Enzyme-linked immunosorbent assay (ELISA), western blotting, immunohistochemistry, and biochemical assays were used to assess the inflammation and the results of signaling pathways associated with ferroptosis. Results:Compared with the control group, the TNBS group had significantly higher DAI scores, histological damage scores, significantly lower protein expression levels of GPX4 and SLC7A11, and higher expression levels of ACSL4 and FTH1. Low- and high-dose TB treatment significantly reduced DAI scores and histological damage scores compared with the TNBS group, with the best treatment effect in the high-dose TB group (all P＜0.05). TB reversed the expression of glutathione and malondialdehyde in intestinal epithelial cells from the TNBS group and up-regulated the GPX4 and SLC7A11 protein expression levels while down-regulating the ACSL4 and FTH1 expression levels (all P＜0.05). Conclusion:TB ameliorates oxidative stress and intestinal inflammatory damage in TNBS-induced Crohn's disease rats. Its mechanism may be related to the inhibition of the ferroptosis pathway.