Bioscience Reports

, Volume 21, Issue 1, pp 33–43 | Cite as

Quantitative Analysis of Phospholipid Peroxidation and Antioxidant Protection in Live Human Epidermal Keratinocytes

  • Anna A. Shvedova
  • Yulia Y. Tyurina
  • Vladimir A. Tyurina
  • Yoko Kikuchi
  • Valerian E. Kagan
  • Peter J. Quinn
Article
  • 8 Downloads

Abstract

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.

keratinocytes phospholipid peroxidation oxidative stress xanthine/xanthine oxidase AMVN, α-tocopherol 

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Copyright information

© Plenum Publishing Corporation 2001

Authors and Affiliations

  • Anna A. Shvedova
    • 1
  • Yulia Y. Tyurina
    • 2
    • 3
  • Vladimir A. Tyurina
    • 2
    • 3
  • Yoko Kikuchi
    • 2
  • Valerian E. Kagan
    • 2
  • Peter J. Quinn
    • 4
  1. 1.Health Effects Laboratory Division, Pathology and Physiology Research BranchNIOSHMorgantown
  2. 2.Department of Environmental and Occupational HealthUniversity of PittsburghPittsburghUSA
  3. 3.Institute of Evolutionary Physiology and BiochemistryRussian Academy of SciencesSt. PetersburgRussia
  4. 4.Division of Life SciencesKing's CollegeLondonUK

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