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The effect of an antioxidative peptide Met-Lys-Ala-Val-Cys-Phe-Ser-Leu (MKAVCFSL) on oxidative stress in Caco-2 cell lines was investigated. Caco-2 cells exposed to excess oxidative stress could be restored when pretreated with the peptide. Reactive oxygen species (ROS) and malondialdehyde (MDA) within the cells could be scavenged by MKAVCFSL. The peptide could also enhance the activity of glutathione peroxidase (GPx), glutathione reductase (GR), and superoxide dismutase (SOD), while catalase (CAT) activity did not show a significant difference between treatment and control samples. Meanwhile, it was observed that peptide treatment increased the concentration of glutathione (GSH). Yet the content of glutathione disulfide (GSSG) was hardly affected. The stability of MKAVCFSL was also assessed and an intact peptide was observed after simulated gastrointestinal digestion. Part of the peptide was hydrolyzed into fragments including MKA, FSL, AVCFSL, and MKAVCF. This study demonstrated that MKAVCFSL derived from bighead carp hydrolysates could ameliorate oxidative stress to protect the Caco-2 cells.