Antioxidant potential and protective effect of modified sea cucumber peptides against H2O2-induced oxidative damage in vitro HepG2 cells and in vivo zebrafish model

Abstract

In this study, modified sea Cucumber Peptides (SCP) were prepared by reacting with xylooligosaccharide (XOS) and alginate oligosaccharides (AOS) via glycation. Free radical inhibitory and inhibition of oxidative stress of modified SCP was evaluated using human hepatocellular carcinoma (HepG2) cells and zebrafish embryos. LC-MS analysis revealed that SCPs mainly consist of 40 active peptides, with an average molecular weight of 1122.168 Da and an average length of 11 amino acid residues. For amino acid composition, L-Asparagine, L-Methionine, and L-Aspartic Acid were dominant amino acids in SCP. The result showed that the antioxidant ability of SCP against 2,2-Diphenyl-1-picrylhydrazyl (DPPH), superoxide anion radical (O^-2), and Hydroxyl Radical (OH) was significantly improved after modification. In HepG2 cells, the modified SCP showed stronger protection than native SCP native against H₂O₂-induced oxidative stress by enhancing cell viability and reducing radical oxygen species (ROS) generation. The inhibition effect of SCP was increased after modification with XOS and AOS by 13 % and 19 % respectively. Further studies displayed that the activity of antioxidative enzymes, including Superoxide dismutase (SOD), Glutathione Peroxidase (GPx), and catalase (CAT), was remarkably enhanced, whereas malondialdehyde (MDA) level was reduced compared with native SCP and H₂O₂-treated groups, thus, improving the intracellular antioxidant defenses. The gene expression analysis showed that the mechanism underlying the modified SCP protective effect may be linked with the capability to regulate Nuclear factor-erythroid factor 2-related factor 2 (NRF2) gene expression. The protective effect of modified SCP against H₂O₂ in vitro was confirmed in vivo by reduced toxicity in zebrafish embryos via improvement of mortality rate, hatching rate, heart beating rate, and deformities of the zebrafish model. However, SCPAOS conjugate displayed greater antioxidant potentials compared to the SCPXOS, the different effects between SCPAOS and SCPXOS could be due to their different antioxidant activity. Thus, modified SCP could be potentially used as a novel nutraceutical in the preparation of anti-aging food and medicine.

Keywords: Antioxidant potential; HepG2 cells; Modification; Oxidative damage; Sea cucumber peptides.