Peptidomics-driven discovery of novel cowhide collagen peptides with in vitro anti-inflammatory activity against ox-LDL-induced macrophage dysfunction

Chronic inflammatory vascular diseases, driven by oxidative stress and lipid dysregulation, are increasingly recognized as the early initiating phenomena of various cardiovascular pathologies. Recent insights highlight animal-derived collagen peptides as beneficial agents for improving vascular function, attributable to their excellent biocompatibility and superior anti-inflammatory and antioxidant effects. This study employed cowhide Alcalase hydrolysate as raw material, utilizing chromatographic purification, peptidomics, molecular simulation, and in vitro assays to screen five bioactive peptides (GPAWR, GPWR, KGPWR, KWCAGPR, LGPRW). CA exhibited complete amino acid profiles and characteristic triple-helix structures, with purified fractions enriched in hydrophobic short peptides. In ox-LDL-induced RAW264.7 cells, these peptides significantly restored nitric oxide levels and suppressed pro-inflammatory cytokines, with LGPRW demonstrating superior efficacy. Under in vitro conditions, these novel bovine dermal collagen peptides effectively ameliorated ox-LDL-induced macrophage dysfunction through NO modulation and inflammation inhibition. These findings provided sequence-level evidence for exploring novel collagen fragments as potential natural anti-inflammatory agents.