Enhanced oxidative stress by L-ascorbic acid within cells challenged by hydrogen peroxide

Journal of Nutritional Science and Vitaminology
Yoko InaiMorimitsu Nishikimi

Abstract

It has been amply documented that L-ascorbic acid added to the medium of a cell culture increases oxidative damage, and this effect of L-ascorbic acid has been ascribed to the generation of reactive oxygen intermediates in the medium during its auto-oxidation. We have here questioned whether such an effect is exerted inside the cell as well, and if so, what its mechanism is. To assess thiol oxidation in the cell, we manipulated CHO cells so that they could express bacterial alkaline phosphatase in the cytoplasm. Alkaline phosphatase activity, which requires the formation of intramolecular disulfide bridges, was shown to appear when the cells were exposed to H2O2. This H2O2-induced activity increased more than 1.5 fold when L-ascorbic acid had been loaded in the cells by incubation with L-ascorbic acid-2-O-phosphate. Similar enhancing effects were also observed by assessing oxidation of glutathione, formation of protein carbonyls, and generation of reactive oxygen intermediates. Interestingly, the effects by the L-ascorbic acid-2-O-phosphate treatment were totally suppressed by addition of the membrane-permeable chelator deferoxamine to the medium, indicating the involvement of iron ions. Because the apoprotein of conalbumin, wh...Continue Reading

Citations

May 5, 2012·The Journal of Nutrition, Health & Aging·W ZhangX Jia
Jul 2, 2021·The Journal of Venomous Animals and Toxins Including Tropical Diseases·Carlos Alberto-SilvaKatsuhiro Konno

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