Investigating the Effects of Different n‐6 Polyunsaturated Fatty Acid Ratios on Ferroptosis in IPEC‐J2 Cells

Dietary n-6 and n-3 polyunsaturated fatty acid (PUFA) balance critically modulates various physiological processes, including inflammation and cell death. This study investigated the effects of different n-6 PUFA ratios (1:1, 5:1, 10:1, 20:1) on ferroptosis in porcine IPEC-J2 intestinal epithelial cells. Cells treated with varying PUFA ratios showed a significant reduction in cell viability, which was alleviated by the ferroptosis inhibitor ferrostatin-1 (fer-1). The 20:1 n-6 PUFA ratio exhibited the most pronounced decrease in cell viability and the greatest increase in malondialdehyde (MDA) levels, a marker of lipid peroxidation. Levels of 4-hydroxynonenal (4-HNE), total iron, and proinflammatory cytokine IL-6 were significantly elevated in the 5:1, 10:1, and 20:1 groups, with fer-1 treatment mitigating these effects. The GSSG/GSH ratio, an indicator of oxidative stress, also significantly increased in the 10:1 and 20:1 groups. These findings suggest that higher n-6 PUFA content exacerbates lipid peroxidation, iron accumulation, and inflammatory responses, leading to ferroptosis in IPEC-J2 cells. This study highlights the importance of dietary n-6 PUFA balance in modulating ferroptosis and intestinal health, offering potential insights into strategies for improving intestinal function by regulating ferroptosis pathways.