Abstract

Oxidative stress can damage lipids, proteins and DNA. The effect of hydrolysed, purified defatted egg yolk protein by-product (fraction EYGF-23) on oxidative stress in Caco-2 colon cancer epithelial cells was investigated for the first time. Caco-2 cells exposed to toxic 3 mM tert-butyl hydroperoxide (t-BHP) for 2 hours were protected when pretreated with 1.0 mg/ml EYGF-23 for 24 hours. The peptides inhibited intracellular reactive oxygen species (ROS) (60.0%) and malondialdehyde (MDA) (24.2 µg/ml) production, compared to cells treated with only t-BHP (75.3% and 32.2 µg/ml respectively). EYGF-23 inhibited caspase 3/7 activity in stressed cells, thereby protecting cells from induced caspase-dependent apoptosis. Further, total glutathione level and superoxide dismutase activity were significantly elevated to 0.29 µM and 55.10% respectively, in cells pretreated with EYGF-23, compared to t-BHP treatment alone (0.10 µM and 44.22 % respectively). Peptides WYGPD, TMFPSA and WIHNENQGF and hydrophobic amino acids tyrosine (Y) and tryptophan (W) contributed to cellular antioxidant activity against oxidative stress in colon epithelial cells and may have nutraceutical applications.

Highlights

  • Reactive oxygen species (ROS) are produced in living cells as a result of normal cell metabolism and xenobiotic detoxification

  • The amount of ROS generated by untreated cells was significantly lower (69.00 %; p

  • ROS production levels in Caco-2 cells treated with 1.0 mg/ml Egg yolk gel filtration fraction-23 (EYGF-23) under oxidative conditions were markedly inhibited to 60.00 % (p < 0.001) when compared with cells treated by t-BHP alone (75.27%)

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Summary

Introduction

Reactive oxygen species (ROS) are produced in living cells as a result of normal cell metabolism and xenobiotic detoxification. Oxidative stress is a consequence of an excess of ROS, which participate in cellular damage by reacting with lipid membranes, proteins, and DNA. ROS are implicated in the pathogenesis of many diseases that include cardiovascular diseases [2] and certain cancers [3] Cellular antioxidative enzymes such as superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) can protect cell membranes and cellular content by scavenging ROS. Previous studies indicated that a phosvitin oligophosphopeptide, isolated from egg yolk protein, protected intestinal cells from oxidative stress induced by hydrogen peroxide by suppressing lipid oxidation, enhancing the synthesis of GSH and increasing its level [9]. The aim of this novel study was to use cultured Caco-2 cells as an epithelial model to investigate the antioxidant effect of isolated egg yolk peptides in cells exposed to pro-oxidant t-BHP

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