Breakage and binding of DNA by reaction products of hypochlorous acid with aniline, 1-naphthylamine, or 1-naphthol.
Hypochlorous acid (HOCl) is a chemically reactive oxidant and a potent microbicidal agent that is synthesized in phagosomes of inflammatory neutrophils and released into extracellular spaces. Besides reducing pathogenicity by reacting with phagocytized infectious agents, HOCl may damage tissues and yield toxic products upon reaction with various other molecules, including xenobiotics. As model xenobiotics, the substituted aryl compounds aniline, 1-naphthylamine, and 1-naphthol (1-NOH) were investigated herein for their potential to react with HOCl and the transformed into genotoxic products. The compounds were first exposed to HOCl (25-150 microM) in phosphate buffer and afterward used to treat human fibroblasts or purified DNA. DNA single-strand breaks in cells and the binding of HOCl-reacted 1-[14C]NOH to purified DNA were assessed by DNA alkaline elution and scintillation spectrometry, respectively. It was found that neither HOCl nor compounds alone could break cellular DNA. But HOCl-reacted compounds produced up to 400 rad equivalents of DNA breaks. HOCl reaction products of aniline and the model bicyclic aryl compounds differed in their DNA-breaking characteristics. HOCl-reacted 1-[14C]NOH was stable and bound to DNA at up to 124 pmol/mg DNA. Sodium thiosulfate, glutathione, and taurine inhibited the transformation reactions; but only the former two blocked binding of HOCl-reacted 1-NOH to DNA. Ultraviolet spectra showed that HOCl reacted rapidly (less than 1 min) and equally well with 1-NOH at pH 7.2 or at an intraphagosomal pH of 5.0. Reaction concentrations of HOCl in this study were 2- to 11-fold lower than levels generated in vitro by stimulated neutrophils. These results show that certain aryl compounds can react readily with approximated physiological levels of HOCl (-OCl) to form relatively long-lived products that bind DNA and are genotoxic to human cells.
Kozumbo
W J
WJ
Center for Environmental Medicine and Lung Biology, University of North Carolina, Chapel Hill 27599.
Agarwal
S
S
Koren
H S
HS
D015057:1-Naphthylamine / Q000737:chemistry / Q000633:toxicity*; D000814:Aniline Compounds / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000633:toxicity*; D016587:Antimutagenic Agents / Q000494:pharmacology; D000975:Antioxidants / Q000494:pharmacology; D002478:Cells, Cultured; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects; D006801:Humans; D006997:Hypochlorous Acid / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000633:toxicity*; D008168:Lung / Q000166:cytology / Q000187:drug effects; D009153:Mutagens / Q000737:chemistry / Q000633:toxicity*; D009284:Naphthols / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000633:toxicity*; D009504:Neutrophils / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D000814:Aniline Compounds; D016587:Antimutagenic Agents; D000975:Antioxidants; D004277:DNA, Single-Stranded; D009153:Mutagens; D009284:Naphthols; C029350:1-naphthol; D006997:Hypochlorous Acid; D015057:1-Naphthylamine; C023650:aniline