Ferrostatin-1 Attenuates CLP-Induced Colonic Injury: Associations with Ferroptosis-Related Alterations, Oxidative Stress, Inflammation, and Treatment Timing

Fecal peritonitis induces severe oxidative and inflammatory injury and may involve ferroptosis-related mechanisms. This study investigated the effects of Ferrostatin-1 (Fer-1) on cecal ligation and puncture (CLP)-induced colonic injury, with emphasis on oxidative stress, inflammation, ferroptosis-related alterations, and treatment timing. Thirty-five male Wistar rats were randomly assigned to five groups: Sham, CLP, CLP + dimethyl sulfoxide (DMSO), CLP + Fer-1 (30 min), and CLP + Fer-1 (2 h). Fer-1 (5 mg/kg, intraperitoneally) was administered after CLP. Biochemical and inflammatory parameters, colonic histopathology, and acyl-CoA synthetase long-chain family member 4 (ACSL4) and glutathione peroxidase 4 (GPX4) immunoreactivity were evaluated. CLP induced marked oxidative and inflammatory alterations, increased histopathological damage and ACSL4 immunoreactivity, and decreased GPX4 immunoreactivity (all overall p < 0.001, except total antioxidant status, p = 0.004). Fer-1 treatment generally shifted these alterations toward the sham profile. In exploratory comparisons adjusted for multiple testing, the 2 h group showed lower malondialdehyde, tumor necrosis factor-alpha, and interleukin-6 levels and higher catalase concentrations than the 30 min group (Holm-adjusted p < 0.05). Group-adjusted analyses indicated that the strong pooled correlations were largely attributable to between-group differences. In silico analyses provided complementary hypothesis-generating molecular context. Fer-1 attenuated CLP-induced colonic injury and was associated with favorable oxidative–inflammatory changes and ferroptosis-related ACSL4/GPX4 alterations. However, the timing findings do not establish an optimal therapeutic window, and direct measures of ferroptotic cell death are required to clarify the contribution of ferroptosis-associated mechanisms.