Transformation and activation of benzidine by oxidants of the inflammatory response.
Aromatic amines, such as benzidine (BZ), initiate bladder cancer in humans. Inflammation/infection play an important role in this cancer. This study was designed to assess the influence of inflammatory oxidants, including reactive nitrogen oxygen species (RNOS), on BZ transformation and activation. RNOS were generated under various conditions and reacted with BZ, and the products were examined by HPLC. Conditions that generate nitrogen dioxide radical, NO(2)(-) + myeloperoxidase + H(2)O(2) and ONOO(-), produced primarily a single new product, which was identified by MS as azobenzidine (AZO-BZ). The myeloperoxidase-catalyzed reaction was inhibited by 1 mM cyanide and did not require NO(2)(-). Chloride (100 mM) reduced the myeloperoxidase reaction by 30% with taurine having little effect. In contrast, conditions that generate N(2)O(3), i.e., NO donor diethylamine (DEA) NONOate, produced two products, which were identified by MS as 4'-OH-4-aminobiphenyl (4'-OH-ABP) and 4-aminobiphenyl (ABP). 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, an oxidant of NO thought to produce NO(2)(*), had a biphasic effect on product formation. At a concentration equal to DEA NONOate, a 5-fold increase in BZ nitrosation was observed, while at higher concentrations nitrosation was greatly diminished and formation of AZO-BZ occurred. Glutathione prevented RNOS transformation of BZ. With MPO and ONOO(-), a new product was formed that cochromatographed with 3-(glutathione-S-yl)BZ. Glutathione also prevented nitrosation of BZ but did not form additional BZ products. HOCl-mediated activation of BZ, 4'-OH-ABP, and ABP to bind DNA was assessed. A higher level of binding was observed at pH 5.5 than pH 7.4. BZ elicited the most binding. More binding was observed at both pH values with 4'-OH-ABP than ABP. Thus, components of the inflammatory response are capable of BZ transformation and activation.
Lakshmi
Vijaya M
VM
Division of Geriatric Medicine, VA Medical Center, St Louis, Missouri 63110, USA.
Hsu
Fong Fu
FF
Zenser
Terry V
TV
D000818:Animals; D001560:Benzidines / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics*; D001565:Benzoates / Q000737:chemistry; D001711:Biotransformation; D002417:Cattle; D004247:DNA / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism; D006997:Hypochlorous Acid / Q000737:chemistry / Q000494:pharmacology; D007093:Imidazoles / Q000737:chemistry; D007249:Inflammation / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D016877:Oxidants / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D026361:Reactive Nitrogen Species / Q000737:chemistry / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000737:chemistry / Q000378:metabolism
D001560:Benzidines; D001565:Benzoates; D007093:Imidazoles; D016877:Oxidants; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; C079393:1,3-dihydroxy-4,4,5,5-tetramethyl-2-(4-carboxyphenyl)tetrahydroimidazole; D006997:Hypochlorous Acid; D004247:DNA; C427203:calf thymus DNA; D006861:Hydrogen Peroxide; D005978:Glutathione