Ferroptosis is an iron-dependent form of regulated cell death induced by the lethal accumulation of lipid peroxidation, while the precise mechanism of ferroptosis in the pathogenesis of ulcerative colitis (UC) remains to be elucidated. This study aimed to explore the potential effect of hypoxia inducible factor-1alpha (HIF-1alpha) on ferroptosis in intestinal epithelial cells (IECs) in UC. The relationship between ferroptosis and HIF-1alpha was initially investigated using clinical UC colon samples. In vitro and in vivo models of acute intestinal inflammatory response were constructed using lipopolysaccharide (LPS) and dextran sulfate sodium (DSS), respectively. The effect of HIF-1alpha on ferroptosis in UC was determined by establishing HIF-1alpha overexpression (HIF1A-OE) or knockdown (shHIF1A) IEC lines, and the mechanism by which HIF-1alpha mediated the transcription of glutathione peroxidase 4 (GPX4) was explored by combining Co-immunoprecipitation (Co-IP) and Chromatin immunoprecipitation-qPCR (ChIP-qPCR). The results indicated that ferroptosis was present in IECs from UC patients and colitis mice. Elevated expression of HIF-1alpha ameliorated the secretion of inflammatory cytokines and ferroptosis in IECs in vitro. HIF-1alpha inhibited ferroptosis by transcriptional activation of the GPX4 gene in inflammatory IECs. HIF-1alpha ameliorated the general conditions of mice and intestinal barrier dysfunction and by suppressing ferroptosis in IECs of mice through upregulating the expression of GPX4. In conclusion, ferroptosis occurred in the IECs of UC patients and colitis mice. HIF-1alpha may improve UC by suppressing ferroptosis in IECs through regulating the transcription of GPX4.