To characterize oxidative stress in phospholipids of normal human epidermal keratinocytes we metabolically labeled their membrane phospholipids with a natural oxidation-sensitive fluorescent fatty acid, cis-parinaric acid, and exposed the cells to two different sources of oxidants–a lipid-soluble azo-initiator of peroxyl radicals, 2,2′-azobis(2,4-dimethyl-valeronitrile), AMVN, and a superoxide generator, xanthine oxidase/xanthine. We demonstrated that both oxidants induced pronounced oxidation of four major classes of cis-parinaric acid-labeled phospholipids–phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and phosphatidylinositol–in normal human epidermal keratinocytes that was not detectable as any significant change of their phospholipid composition. Vitamin E was effective in protecting the cells against phospholipid peroxidation. Since viability of normal human epidermal keratinocytes was not changed either by labeling or exposure to oxidants the labeling protocol and oxidative stress employed are compatible with the quantitative analysis of phospholipid peroxidation in viable cells.
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February 01 2001
Quantitative Analysis of Phospholipid Peroxidation and Antioxidant Protection in Live Human Epidermal Keratinocytes
Anna A. Shvedova;
Anna A. Shvedova
1Health Effects Laboratory Division, Pathology and Physiology Research Branch, NIOSH, 1095 Willowdale Road, Morgantown, WV 26505
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Yulia Y. Tyurina;
Yulia Y. Tyurina
2Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15238, USA
3Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, 194223 Russia
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Vladimir A. Tyurina;
Vladimir A. Tyurina
2Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15238, USA
3Institute of Evolutionary Physiology and Biochemistry, Russian Academy of Sciences, St. Petersburg, 194223 Russia
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Yoko Kikuchi;
Yoko Kikuchi
2Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15238, USA
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Valerian E. Kagan;
Valerian E. Kagan
2Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15238, USA
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Peter J. Quinn
Peter J. Quinn
4Division of Life Sciences, King's College, London, UK
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Publisher: Portland Press Ltd
Online ISSN: 1573-4935
Print ISSN: 0144-8463
© 2001 Plenum Publishing Corporation
2001
Biosci Rep (2001) 21 (1): 33–43.
Citation
Anna A. Shvedova, Yulia Y. Tyurina, Vladimir A. Tyurina, Yoko Kikuchi, Valerian E. Kagan, Peter J. Quinn; Quantitative Analysis of Phospholipid Peroxidation and Antioxidant Protection in Live Human Epidermal Keratinocytes. Biosci Rep 1 February 2001; 21 (1): 33–43. doi: https://doi.org/10.1023/A:1010430000701
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