Anti‐Inflammatory Effects of Hen Egg White Hydrolysate and Its Specific Peptides IS8, PA11, and PK8 on LPS‐Induced Macrophage Inflammation

Egg white hydrolysate (EWH) has garnered significant attention for the therapeutic properties of their bioactive peptides. This study provides a comprehensive peptidomics analysis of EWH, focusing on its anti‐inflammatory potential. Peptides were classified based on their anti‐inflammatory potential (AIP) scores to identify promising candidates. In LPS‐stimulated RAW 264.7 macrophages, specific peptides effectively suppressed nitric oxide (NO) production, a key inflammatory marker. IS8, PA11, and PK8 were identified as the most promising candidates. qRT‐PCR analysis revealed their mechanism of action through modulation of key cytokines, including TNF‐α, IL‐1β, and IL‐10, as well as other mediators such as iNOS and COX‐2. Molecular docking studies provided additional insights into the anti‐inflammatory potential of these peptides. Notably, IS8, PA11, and PK8 exhibited strong binding affinities to the iNOS active site, with IS8 and PK8 achieving the highest docking scores. The peptides engaged in critical interactions within the iNOS binding pocket through hydrogen bonding and π‐π stacking with key residues, suggesting their potential to inhibit iNOS catalytic activity and thereby reduce NO production. These findings highlight the therapeutic potential of EWH‐derived peptides, particularly IS8, PA11, and PK8, as anti‐inflammatory agents through their ability to modulate key inflammatory pathways, with iNOS inhibition as a central mechanism.