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Journal of Histochemistry and Cytochemistry, Vol. 50, 341-351, March 2002, Copyright © 2002, The Histochemical Society, Inc.

ARTICLE

Evidence of Oxidative Stress in Human Corneal Diseases

Rajeev Buddib, Brian Lina, Shari R. Atilanoa, Nadia C. Zorapapela, M. Cristina Kenneya, and Donald J. Browna
a Ophthalmology Research Laboratories, Department of Surgery, Burns & Allen Research Institute, Cedars–Sinai Medical Center, University of California at Los Angeles Medical School Affiliate, Los Angeles, California
b The Eye Institute, Medical College of Wisconsin, Milwaukee, Wisconsin

Correspondence to: Donald J. Brown, Ophthalmology Research, Cedars–Sinai Medical Center, D-5069, 8700 Beverly Blvd., Los Angeles, CA 90048. E-mail: brown2@cshs.org

This study localized malondialdehyde (MDA, a toxic byproduct of lipid peroxidation), nitrotyrosine [NT, a cytotoxic byproduct of nitric oxide (NO)], and nitric oxide synthase isomers (NOS) in normal and diseased human corneas. Normal corneas (n=11) and those with clinical and histopathological diagnoses of keratoconus (n=26), bullous keratopathy (n=17), and Fuchs' endothelial dystrophy (n=12) were examined with antibodies specific for MDA, NT, eNOS (constitutive NOS), and iNOS (inducible NOS). Normal corneas showed little or no staining for MDA, NT, or iNOS, whereas eNOS was detected in the epithelium and endothelium. MDA was present in all disease groups, with each group displaying a distinct pattern of staining. NT was detected in all keratoconus and approximately one half of Fuchs' dystrophy corneas. iNOS and eNOS were evident in all the diseased corneas. Keratoconus corneas showed evidence of oxidative damage from cytotoxic byproducts generated by lipid peroxidation and the NO pathway. Bullous keratopathy corneas displayed byproducts of lipid peroxidation but not peroxynitrite (MDA but not NT). Conversely, Fuchs' dystrophy corneas displayed byproducts of peroxynitrite with little lipid peroxidation (NT >> MDA). These data suggest that oxidative damage occurs within each group of diseased corneas. However, each disease exhibits a distinctive profile, with only keratoconus showing prominent staining for both nitrotyrosine and MDA. These results suggest that keratoconus corneas do not process reactive oxygen species in a normal manner, which may play a major role in the pathogenesis of this disease.

(J Histochem Cytochem 50:341–351, 2002)

Key Words: peroxynitrite, nitric oxide, nitrotyrosine, keratoconus, bullous keratopathy, Fuchs' endothelial dystrophy, malondialdehyde, lipid peroxidation


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