I3C protects IPEC-J2 cells by inhibiting ferroptosis through activation of the AhR-Nrf2-SLC7A11-GPX4 pathways

The intestinal epithelial barrier function in piglets can be impaired by weaning stress. Indole-3-carbinol (I3C) has been reported to alleviate intestinal barrier damage in piglets through aryl hydrocarbon receptor (AhR) activation, but the underlying mechanisms remain unclear. This study aimed to investigate the molecular mechanisms by which I3C-activated AhR regulates the Nrf2-SLC7A11-GPX4 signaling pathway to mitigate barrier dysfunction and ferroptosis in IPEC-J2 cells. It has been observed that the addition of I3C can alleviate LPS-induced damage. Concurrently, a significant upregulation in the gene expression of both the AhR pathway and ferroptosis-related pathways was identified by transcriptomic sequencing. Based on KEGG analysis, it was further speculated that this phenomenon is likely achieved through the ferroptosis pathway. When ferroptosis was inhibited, a reduction in inflammatory response levels, enhanced antioxidant capacity, and increased tight junction formation were detected in IPEC-J2 cells. This protective effect is hypothesized to be mediated through the AhR and Nrf2 signaling pathways. After inhibiting AhR and Nrf2 pathways, the protective effects of I3C on IPEC-J2 cells were reversed, leading to barrier dysfunction and a significant increase in ferroptosis markers. In conclusion, it was demonstrated that I3C alleviates LPS-induced intestinal barrier injury in IPEC-J2 cells through AhR-mediated activation of the Nrf2-SLC7A11-GPX4 signaling pathway, thereby suppressing ferroptosis.