PSI-16 Intestinal Porcine Enterocyte Cell (Ipec-J2) as an In-Vitro Model for Investigating the Mechanisms of Ferroptosis

Abstract

Abstract Ferroptosis is a newly identified form of regulated cell death that has been implicated in various intestinal diseases. However, there is limited research on ferroptosis in porcine models. The aim of this study was to establish an in vitro model of ferroptosis in the IPEC-J2 cell line. Ferroptosis is a form of regulated cell death that was first identified in 2012 and is characterized by unique morphological and physiological features that distinguish it from other types of cell death. This process is triggered by a complex interplay of multiple factors, including excess ferrous iron accumulation, oxidation of polyunsaturated fatty acids, and dysregulation of redox reactions. These events ultimately lead to the accumulation of lipid peroxides and subsequent membrane damage, resulting in ferroptosis. In pig production, weaning piglets often experience significant stress that can cause damage to the intestinal barrier due to changes in diet and environment, leading to diarrhea and growth retardation. Whether ferroptosis is involved in the development of intestinal diseases in pigs is still unclear, this study will establish a ferroptosis model using porcine intestinal epithelial cells. To induce ferroptosis in the IPEC-J2 cells, we applied hydrogen peroxide (H2O2), lipopolysaccharide (LPS), ferrous sulfate (FeSO4), and erastin, a ferroptosis inducer. The concentration of each treatment was determined by analyzing cell viability using the CCK-8 assay. Lipid peroxidation was evaluated using the TBARS assay, and the data indicated that compared with the control samples, IPEC-J2 cells treated with H2O2, FeSO4 and erastin had greater malondialdehyde levels (fold of control; CON = 1, H2O2 = 1.70, FeSO4 = 2.32, erastin = 1.68; P <0.05). Iron concentrations were measured using the Iron Assay kit (cat. no. ab83366; Abcam). Greater total iron concentrations were observed in IPEC-J2 cells treated with FeSO4 and erastin (fold of control; CON = 1, FeSO4 = 14.91, erastin = 1.10; P <0.05), and higher ferrous iron level on IPEC-J2 cells treated with erastin (fold of control; CON = 1, erastin = 1.17; P <0.05). In conclusion, the above results suggest that treatment with H2O2, FeSO4 and erastin is likely to lead to ferroptosis. However, further investigation is needed to confirm these findings. A ferroptosis model on intestinal porcine epithelial cells will be established in this study.