Anti-inflammatory effect of two novel peptides derived from Binglangjiang buffalo whey protein in lipopolysaccharide-stimulated RAW264.7 macrophages

Whey protein hydrolysate from Binglangjiang buffalo, a unique genetic resource, has anti-inflammatory activity, but its anti-inflammatory composition and effects are unknown. The aim of this study was to investigate the anti-inflammatory peptides from Binglangjiang buffalo whey protein hydrolysate. A total of 1483 peptides were identified using LC-MS/MS, and 12 peptides were chosen for chemical synthesis using peptidomics, and then two novel anti-inflammatory peptides (DQPFFHYN (DN8) and YSPFSSFPR (YR9)) were screened out using LPS-stimulated RAW264.7 cells. The molecular weights of DN8 and YR9 with β-turn conformations were 1067.458 Da and 1087.52 Da, respectively, and showed a high in-vitro safety profile and thermal stability, but were intolerant to pepsin. Furthermore, ELISA and Western blot analysis indicated that peptides DN8 and YR9 significantly suppressed the secretion of pro-inflammatory cytokines NO, TNF-α, and IL-6 and the expression of mediators iNOS, TNF-α, and IL-6 in LPS-stimulated RAW264.7 cells. The study provides insights into the development of novel food-based anti-inflammatory nutritional supplements.