Xanthine oxidoreductase and xanthine oxidase in human cornea.
Xanthine oxidoreductase (xanthine dehydrogenase + xanthine oxidase) is a complex enzyme that catalyzes the oxidation of hypoxanthine to xanthine, subsequently producing uric acid. The enzyme complex exists in separate but interconvertible forms, xanthine dehydrogenase and xanthine oxidase, which generate reactive oxygen species (ROS), a well known causative factor in ischemia/reperfusion injury and also in some other pathological states and diseases. Because the enzymes had not been localized in human corneas until now, the aim of this study was to detect xanthine oxidoreductase and xanthine oxidase in the corneas of normal post-mortem human eyes using histochemical and immunohistochemical methods. Xanthine oxidoreductase activity was demonstrated by the tetrazolium salt reduction method and xanthine oxidase activity was detected by methods based on cerium ion capture of hydrogen peroxide. For immunohistochemical studies. we used rabbit antibovine xanthine oxidase antibody, rabbit antihuman xanthine oxidase antibody and monoclonal mouse antihuman xanthine oxidase/xanthine dehydrogenase/aldehyde oxidase antibody. The results show that the enzymes are present in the corneal epithelium and endothelium. The activity of xanthine oxidoreductase is higher than that of xanthine oxidase, as clearly seen in the epithelium. Further studies are necessary to elucidate the role of these enzymes in the diseased human cornea. Based on the findings obtained in this study (xanthine oxidoreductase/xanthine oxidase activities are present in normal human corneas), we hypothesize that during various pathological states, xanthine oxidase-generated ROS might be involved in oxidative eye injury.
Cejková
J
J
Department of Eye Histochemistry, Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Prague. cejkova@biomed.cas.cz
Ardan
T
T
Filipec
M
M
Midelfart
A
A
D000328:Adult; D003315:Cornea / Q000201:enzymology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007150:Immunohistochemistry; D008875:Middle Aged; D017382:Reactive Oxygen Species; D014968:Xanthine Dehydrogenase / Q000096:biosynthesis*; D014969:Xanthine Oxidase / Q000096:biosynthesis* / Q000378:metabolism
D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase