黄鳝养殖废液中活性肽筛选及其抗氧化应激与皮肤愈合活性OACHSSCD

[Objective]This study aims to explore the utilization value of bioactive peptides derived from the aquaculture wastewater of Asian swamp eel(Monopterus albus),screen candidate peptides with antioxidant and cell repair functions,and provide a theoretical basis for their application in novel biomaterials.[Methods]Using eel aquaculture wastewater as raw material,peptide mixtures were prepared via physical enrichment and pepsin hydrolysis.Candidate peptides were screened based on bioactivity,water solubility,toxicity and allergenicity,and their binding modes with the Keap1 protein were evaluated via molecular docking.A lipopolysaccharide(LPS)-induced oxidative stress model was established using mouse monocyte macrophages(RAW264.7)to detect the intracellular levels of reactive oxygen species(ROS),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA).Furthermore,a scratch wound model in human skin fibroblasts(HSF)was utilized to evaluate the effects of the active peptides on cell migration.[Results]The mass fraction of protein in the lyophilized powder of the wastewater was 3.12%.The enzymatic hydrolysates were mainly small-molecule peptides with 4 to 15 amino acid residues,which were rich in hydrophobic and partially aromatic amino acids.Five non-toxic candidate wastewater-derived peptides with good water solubility were obtained through multi-dimensional screening,among which the sequence FEGPPFKFKFTLR(hereafter referred to as FEG-13)exhibited the lowest binding free energy with Keap1 and formed a stable complex conformation.Cytological evaluation demonstrated that FEG-13 intervention could significantly decrease the intracellular ROS level in LPS-induced RAW264.7 cells by 48.7%(P<0.001),significantly reduce the MDA molality to 5.23 nmol/mg(P<0.01),and significantly restore the activities of SOD and GSH-Px to 12.77 and 23.12 U/mg(P<0.01),respectively.Meanwhile,intervention with 1.0 mg/mL FEG-13 significantly enhanced the scratch closure rate of HSF cells to 64.43%(P<0.001).[Conclusion]Eel aquaculture wastewater holds potential to obtain various bioactive peptides.The obtained FEG-13 can alleviate cellular oxidative stress by regulating the Keap1-Nrf2 pathway and drive fibroblast migration,exhibiting outstanding potential for antioxidant activity and cell repair.