Comparative transcriptomic analysis reveals the protective mechanism of ferulic acid against lipopolysaccharide-induced ferroptosis in IPEC-J2 cells

Lipopolysaccharide (LPS) is a key virulence factor that induces intestinal oxidative damage in livestock and poultry, and cellular ferroptosis is one of the triggers of oxidative stress. Ferulic acid (FA) is a natural phenolic acid with antioxidant, anti-inflammatory and anti-ferroptosis properties. This study aimed to investigate the protective effects of FA against LPS-induced ferroptosis in the porcine small intestinal epithelial cell line (IPEC-J2) and to elucidate the underlying mechanisms. The experiment included four treatment groups: control group (CON), 50 μg/mL FA group (FA), 10 μg/mL LPS group (LPS), and 50 μg/mL FA combined with 10 μg/mL LPS group (FA and LPS), with all treatments lasting 24 h. The results showed that FA inhibited ferroptosis in IPEC-J2 cells. LPS successfully induced ferroptosis, as evidenced by decreased cell viability, increased levels of ROS, MDA, and Fe 2+ , downregulation of GPX4 and SLC7A11 , and upregulation of TFRC and ACSL4 . FA effectively alleviated LPS-induced ferroptosis in IPEC-J2 cells, with an efficacy comparable to that of the ferroptosis inhibitor Fer-1. Transcriptomic analysis revealed that FA significantly reversed the gene expression changes induced by LPS, with differentially expressed genes enriched in pathways related to ferroptosis, TNF signaling, NF-κB signaling, arachidonic acid metabolism, and PI3K-Akt signaling. Protein–protein interaction (PPI) network analysis indicated that genes such as HMGCS1 , CXCL8 , and IL-18 may serve as key nodes mediating the protective effects of FA. In conclusion, FA alleviates LPS-induced ferroptosis in IPEC-J2 cells by regulating glutathione metabolism, maintaining iron homeostasis, and inhibiting lipid peroxidation. This effect is closely associated with the coordinated regulation of inflammatory responses, lipid metabolism, and ferroptosis-related signaling pathways.