The Characterization and Identification of Cyperus Protein: An In Vitro Study on Its Antioxidant and Anti-Inflammatory Potential

<b>Background:</b> Oxidative stress and inflammation are major drivers of metabolic inflammatory diseases, and natural antioxidant peptides represent promising therapeutic agents. Antioxidant peptides derived from Cyperus protein (CAOP) exhibit high digestibility and bioavailability, but their antioxidant and anti-inflammatory mechanisms remain unclear. <b>Methods:</b> We employed in vitro experiments, non-targeted metabolomics, peptide omics, and molecular docking techniques to explore how CAOP exerts dual antioxidant and anti-inflammatory effects. <b>Results:</b> The in vitro experiments showed that in LPS-induced RAW264.7 cells, CAOP not only significantly increased the levels of superoxide dismutase (SOD) and catalase (CAT) but also significantly reduced the gene expression and secretion of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), as well as the phagocytic ability of cells. Metabolomics studies indicate that CAOP protects cells from LPS-induced damage by enhancing intracellular glutathione metabolism pathways, glyceraldehyde and dicarboxylic acid metabolism pathways, pantothenic acid and coenzyme A biosynthesis metabolism pathways, and thiamine metabolism pathways while inhibiting the ferroptosis pathway. CAOP was purified using Sephadex G-25 column chromatography, and its amino acid sequence was determined using LC-MS/MS technology. Subsequently, 25 peptide sequences were screened through bioinformatics analysis. These peptides can target Keap1. Among them, DLHMFVWS (-ICE = 62.8072) and LGHPWGNAPG (-ICE = 57.4345) are most likely to activate the Nrf2-Keap1 pathway. <b>Conclusions:</b> CAOP exerts antioxidant and anti-inflammatory effects by regulating the key metabolic networks, demonstrating its therapeutic promise for associated with oxidative damage and metabolic inflammation disorders.