Journal
BIOSCIENCE REPORTS
Volume 21, Issue 1, Pages 33-43Publisher
KLUWER ACADEMIC/PLENUM PUBL
DOI: 10.1023/A:1010430000701
Keywords
keratinocytes; phospholipid peroxidation; oxidative stress; xanthine/xanthine oxidase; AMVN; alpha-tocopherol
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Funding
- NINDS NIH HHS [F05 NS 10669] Funding Source: Medline
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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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