Document Detail


Lack of antioxidative properties of vitamin C and pyruvate in cultured retinal pigment epithelial cells.
MedLine Citation:
PMID:  16868779     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Oxidative damage to the retinal pigment epithelium might be involved in the pathogenesis of age related macular degeneration. Thus antioxidative protection represents a rationale for a causative therapy or prophylaxis. The aim of the present study is to evaluate antioxidative properties of vitamin C and pyruvate at retinal pigment epithelial (RPE) cells exposed to oxidative stress. METHODS: The ability of vitamin C and pyruvate to quench hydroxyl radicals was tested using the di-hydro-rhodamine (DHR) assay. Cells of the human RPE cell line ARPE-19 were exposed for 8 min to hydroxyl radicals generated by the Fenton reaction from 2.25 mM H2O2 and 30 microM Fe3+ -nitrilo-tri-acetate. This was done in the absence and presence of 0.3-3.0 mM pyruvate and vitamin C, respectively. Cell survival was analysed by vitality staining (life-dead-assay) and expressed as cell survival ratio. A survival ratio <1.0 indicates cell loss. RESULTS: At concentrations from 0.1 to 1.0 mM vitamin C and pyruvate quench hydroxyl radicals in the DHR assay in absence of living matter. In the presence of 0.1- 0.3 mM vitamin C and pyruvate, ARPE-19 showed a reduced survival ratio (0.87 +/- 0.01 to 0.89 +/- 0.02 after 6 h) which was not the case at the higher concentrations between 1 and 3 mM. The exposure of ARPE-19 cells to hydroxyl radicals reduced the survival ratio to 0.92 +/- 0.02. At concentrations at which vitamin C and pyruvate exert toxic effects, a potentiation of radical induced cell death can be observed (survival ratio 0.79 +/- 0.02 and 0.82 +/- 0.03, respectively). Higher concentrations of vitamin C or pyruvate had no explicit protective effect to the hydroxyl radical induced damage. DISCUSSION: Although vitamin C and pyruvate are potent hydroxyl radical quenchers in vitro they failed to protect cultured ARPE-19 cells from oxidative stress induced cell death. In contrast, when applying the scavengers at low concentrations a potentiation of cell damage was observed.
Authors:
Oliver Zeitz; Lars Schlichting; Gisbert Richard; Olaf Strauss
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv für klinische und experimentelle Ophthalmologie     Volume:  245     ISSN:  0721-832X     ISO Abbreviation:  Graefes Arch. Clin. Exp. Ophthalmol.     Publication Date:  2007 Feb 
Date Detail:
Created Date:  2007-10-23     Completed Date:  2007-12-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8205248     Medline TA:  Graefes Arch Clin Exp Ophthalmol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  276-81     Citation Subset:  IM    
Affiliation:
Universitätsklinikum Hamburg-Eppendorf, Klinik und Poliklinik ffir Augenheilkunde, Martinistr. 52, 20246 Hamburg, Germany. zeitz@uke.uni-hamburg.de
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MeSH Terms
Descriptor/Qualifier:
Antioxidants / pharmacology*
Ascorbic Acid / pharmacology*
Cell Survival
Cells, Cultured
Free Radical Scavengers / pharmacology
Humans
Hydrogen Peroxide / toxicity
Hydroxyl Radical
Oxidative Stress / drug effects
Pigment Epithelium of Eye / cytology,  drug effects*
Pyruvic Acid / pharmacology*
Chemical
Reg. No./Substance:
0/Antioxidants; 0/Free Radical Scavengers; 127-17-3/Pyruvic Acid; 3352-57-6/Hydroxyl Radical; 50-81-7/Ascorbic Acid; 7722-84-1/Hydrogen Peroxide

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