Elsevier

Experimental Eye Research

Volume 49, Issue 4, October 1989, Pages 685-698
Experimental Eye Research

Repair of H2O2-induced DNA damage in bovine lens epithelial cell cultures

https://doi.org/10.1016/S0014-4835(89)80063-6Get rights and content

H2O2 concentrations only slightly higher than normal physiological levels found in the lens andaqueous fluid produce a significant number of DNA single-strand breaks in lens epithelial cell cultures. In this investigation, the repair of DNA damaged by short-term, H2O2-induced oxidation was examined in bovine lens epithelial cell cultures. Repair was rapidly initiated and was almost completed in 30 min. A drop in NAD concentration was associated with the DNA damage. 3-Aminobenzamide inhibition of poly(ADP-ribose) polymerase, an enzyme believed to be stimulated by DNA oxidation and involved in DNA repair, prevented the loss of NAD. In contrast, a similar drop in ATP concentration was only slightly lessened by the presence of this inhibitor. Inhibition of the polymerase by 3-aminobenzamide primarily affected only the early recovery period. Overall, recovery occurred almost as effectively in the presence of the inhibitor as in its absence.

Preincubation of lens cultures with o-phenanthroline, an iron chelator, prevented the drop inNAD levels associated with DNA damage. Since a hydroxyl radical is produced from H2O2 by a Fenton type reaction, this result supports the concept that the H2O2-induced oxidation of DNA is caused by hydroxyl radical. In contrast, peroxide-induced loss of activity of a cytosolic enzyme, glyceraldehyde-3-phosphate dehydrogenase, was unaffected by the presence of o-phenanthroline, suggesting direct H2O2 oxidation of this enzyme.

The results of these experiments suggest that lens epithelium contains enzymes that rapidlyrepair single-strand DNA breaks induced by H2O2 insult.

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    Recipient of a National Institutes of Health Individual National Research Service Award.

    Current address: Department of Molecular Pharmacology, Merck, Sharp and Dohme Research Laboratories, Rahway, NJ07065, U.S.A.

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