A novel rapeseed-derived peptide FHLY alleviates deoxynivalenol-induced enterotoxicity by inhibiting the TLR4/NF-κB/NLRP3 pathway

The common mycotoxin deoxynivalenol (DON) impairs intestinal health, prompting interest in natural, food-based detoxification strategies. This study aimed to identify and evaluate bioactive peptides from rapeseed protein hydrolysate (RPH) capable of alleviating DON-induced enterotoxicity. RPH was extracted from defatted rapeseed meal and demonstrated efficacy against DON-induced enterotoxicity in vivo. Molecular characterization revealed the <1 kDa fraction as predominant (76.09 %), with subsequent peptide sequencing. Bioinformatics and network pharmacology identified FHLY, FYSI, and YYF as promising candidates. Synthesized peptides were evaluated in DON-exposed intestinal epithelial cells. FHLY significantly mitigated enterotoxicity in IPEC-J2 cells by inhibiting the TLR4/NF-κB/NLRP3 signaling pathway. Our findings highlight FHLY as a promising functional food ingredient against mycotoxin toxicity and establish a discovery pipeline for identifying bioactive peptides that exhibit therapeutic effects comparable to interventions targeting DON-induced enterotoxicity. Future research will focus on validating the in vivo efficacy of FHLY and exploring its application in functional foods and feeds. • Rapeseed protein hydrolysates mitigate the intestinal damage associated with deoxynivalenol exposure in vivo models. • FHLY, FYSI, and YYF were identified from RPH as promising candidates to alleviate deoxynivalenol-induced intestinal damage. • FHLY mitigates DON-induced enterotoxicity by inhibiting the TLR4/NF-κB/NLRP3 pathway in IPEC-J2 cells.