Modelling ferroptosis in IPEC-J2 cells: insights into iron-dependent cell death

Ferroptosis, a form of regulated cell death characterised by iron accumulation and lipid peroxidation, plays a crucial role in various diseases. However, its mechanisms in livestock, particularly in pigs, remain unclear. In this study, we established an <i>in vitro</i> ferroptosis model using the porcine intestinal epithelial cell line IPEC-J2 to investigate ferroptosis mechanisms in the intestinal epithelium. IPEC-J2 cells were treated with hydrogen peroxide (H₂O₂), ferrous sulphate (FeSO₄), and the ferroptosis inducer erastin. Our results demonstrated that FeSO₄ successfully induced ferroptosis, as evidenced by increased lipid peroxidation markers, such as malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE). In the case of erastin, ferroptosis induction was confirmed by the increase in MDA levels, and the differential regulation of ferroptosis-related genes <i>ACSL4</i>, <i>TFR1</i>, and <i>FTH1</i> in response to FeSO₄ and erastin treatments. Co-treatment with the ferroptosis inhibitor ferrostatin-1 (fer-1) alleviated ferroptosis-induced lipid peroxidation and reduced cell death, further confirming the occurrence of ferroptosis. Conversely, H₂O₂ treatment increased oxidative stress without inducing ferroptosis-specific markers, suggesting that H₂O₂ does not trigger ferroptosis in IPEC-J2 cells. This study establishes a robust ferroptosis model in porcine intestinal epithelial cells and provides insights into the role of ferroptosis in intestinal health, offering a valuable platform for further research in the context of livestock. FeSO₄ and erastin treatments significantly increased lipid peroxidation markers and ferroptosis-related gene expressionCo-treatment with ferrostatin-1 effectively reduced ferroptosis-induced lipid peroxidation and cell death.H₂O₂ increased oxidative stress but did not induce ferroptosis-specific markers in IPEC-J2 cells. FeSO₄ and erastin treatments significantly increased lipid peroxidation markers and ferroptosis-related gene expression Co-treatment with ferrostatin-1 effectively reduced ferroptosis-induced lipid peroxidation and cell death. H₂O₂ increased oxidative stress but did not induce ferroptosis-specific markers in IPEC-J2 cells.