AIM: To investigate the effect of autophagy on cell ferroptosis in intestinal ischemia-reperfusion injury. METHODS: Twenty-four SPF grade Wistar rats weighing 200-220 g were divided into 4 groups (n=6): sham operation group (sham group), ischemia group (I group), ischemia-reperfusion group (I/R group),and ischemia-reperfusion+autophagy inhibitor group (I/R+3-MA group).The ischemia model was established by clamping the superior mesenteric artery for 1 hour, and the intestinal ischemia-reperfusion injury model was established by reperfusion for 2 hours. HE staining was used to observe the pathological changes of intestinal mucosa and Chiu score under light microscope. Fe2+ contents was measured by Colorimetric and LDH, LPO contents were measured by ELISA. FTH1, NCOA4, and LC3II/I expression by WB, and mitochondrial morphology was measured by transmission electron microscopy. RESULTS: Compared with the sham group, the remaining three groups had higher Chiu scores, FTH1 and NCOA4 were downregulated and LC3II/I was upregulated (P<0.01), LDH and Fe2+ were increased (P<0.01) in I and I/R, LDH (P<0.05), Fe2+ (P<0.01) were increased in I/R+3-MA group, the LPO content was elevated (P<0.01) in I/R; Compared with group I, the Chiu score (P<0.01), LDH, Fe2+ (P<0.05) and LPO (P<0.01) content were increased in group I/R (P<0.01), FTH1, NCOA4 was downregulated and LC3II/I was upregulated (P<0.01); Compared with the I/R, The Chiu score was decreased, the LDHand LPO (P<0.01), Fe2+ (P<0.05) content were decreased in I/R+3-MA, FTH1, NCOA4 was up-regulated and LC3II/I was down-regulated (P<0.01). Mitochondrial morphological changes: in sham group, intestinal tissue and mitochondria were intact, I group the mitochondrial cristae is reduced, and the double membrane structure is incomplete; in I/R group, the mitochondrial cristae was greatly reduced, and the organelle damage was serious. Increincrease in mitochondrial number and internal cristae were observed after 3-MA inhibition, and the membrane gradually became intact. CONCLUSION: In intestinal ischemia-reperfusion injury, autophagy participates in the regulation of cellular iron ions and promotes the occurrence of cell ferroptosis, and the inhibition of autophagy can reduce I/R intestinal injury.