The xenobiotic-metabolizing enzymes arylamine N-acetyltransferases in human lens epithelial cells: inactivation by cellular oxidants and UVB-induced oxidative stress.
The human arylamine N-acetyltransferases NAT1 and NAT2 are important xenobiotic-metabolizing enzymes involved in the detoxification and metabolic activation of numerous drugs and chemicals. NAT activity depends on genetic polymorphisms and on environmental factors. It has been shown that low NAT-acetylation activity could increase the risk of age-dependent cataract, suggesting that NAT detoxification function may be important for lens cells homeostasis. We report here that the NAT acetylation pathway may occur in human lens epithelial (HLE) cells. Functional NAT1 enzyme was readily detected in HLE cells by reverse transcription-polymerase chain reaction, Western blotting, and enzyme activity assays. NAT2 mRNA and enzymic activity were also detected. We investigated whether oxidants, known to be produced in HLE cells during oxidative stresses and involved in age-dependent cataract formation, decreased endogenous NAT1 and NAT2 activity. The exposure of HLE cells to peroxynitrite led to the dose-dependent irreversible inactivation of both NAT isoforms. Exposing HLE cells to continuously generated H(2)O(2) gave a dose-dependent inactivation of NAT1 and NAT2, reversible on addition of high concentrations of reducing agents. UVB irradiation also induced the reversible dose-dependent inactivation of endogenous NAT1 and NAT2, reversible on addition of reducing agents. Thus, our data suggest that functional NAT1 and NAT2 are present in HLE cells and may be impaired by oxidants produced during oxidative and photooxidative stresses. Oxidative-dependent inhibition of NATs in these cells may increase exposure of lens to the harmful effects of toxic chemicals that could contribute to cataractogenesis over time.
Dairou
Julien
J
Centre National de la Recherche Scientifique-Unité Mixte de Recherche 7000, Faculté de Médecine Pitié-Salpêtrière, Paris, France.
Malecaze
François
F
Dupret
Jean-Marie
JM
Rodrigues-Lima
Fernando
F
D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000378:metabolism; D002386:Cataract / Q000209:etiology*; D002460:Cell Line; D004847:Epithelial Cells / Q000201:enzymology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007527:Isoenzymes; D007908:Lens, Crystalline / Q000201:enzymology*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D014466:Ultraviolet Rays
D007527:Isoenzymes; D016877:Oxidants; D006861:Hydrogen Peroxide; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; C478900:NAT2 protein, human