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 in vitro 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 ACSL4, TFR1, and FTH1 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.