title: Biochemical responses of skin to allergenic and non-allergenic nitrohalobenzenes. Evidence that an NADPH-dependent reductase in skin may act as a prohapten-activating enzyme.
abstract: Using a selection of 'classic' haptens (dinitrohalobenzenes and picryl chloride) and related non-sensitizing analogous, we examined changes in levels of glutathione (GSH) and glutathione disulphide (GSSG) in mouse skin 12 h after their epicutaneous application. We observed that elevation of GSSG levels and/or depletion of GSH levels correlated well with contact allergenic potential. Non-sensitizing analogous failed to perturb GSH/GSSG status. In vitro assays using mouse skin and rat liver microsomal preparations indicated that only the allergenic nitrohalobenzenes initiated NADPH-dependent oxygen utilization, with the activity falling off in the order picryl chloride >> DNIB > DNBB > DNCB > DNFB. In addition, an examination of the colour of mouse skin homogenates ex vivo after application of the dinitrohalobenzenes showed significant yellowing (consistent with aromatic nucleophilic substitution) only with DNFB. Our results indicate that, while an aromatic nucleophilic substitution reaction with skin protein can possibly account for the allergenicity of DNFB, it does not seem to occur with DNCB, DNBB or DNIB. These may instead behave mainly as prohaptens which are activated enzymically by NADPH-dependent reductase(s) within the skin, with the concomitant generation of superoxide and hydrogen peroxide, to form potentially protein-reactive free radical and other metabolites. Picryl chloride appears capable of both conjugating directly with proteins by aromatic nucleophilic substitution and undergoing NADPH-dependent metabolism to other potentially protein-reactive metabolites.
authors.affiliation: Welsh School of Pharmacy, University of Wales College of Cardiff, UK.
authors.forename: R J
authors.initials: RJ
authors.lastname: Schmidt
authors.forename: L Y
authors.initials: LY
authors.lastname: Chung
mesh_terms: D000818:Animals; D017449:Dermatitis, Allergic Contact / Q000201:enzymology / Q000209:etiology*; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism; D019803:Glutathione Disulfide; D006846:Hydrocarbons, Halogenated / Q000633:toxicity*; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009247:NADH, NADPH Oxidoreductases / Q000187:drug effects*; D009249:NADP / Q000378:metabolism; D009578:Nitrobenzenes / Q000633:toxicity*; D010100:Oxygen / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012867:Skin / Q000187:drug effects* / Q000201:enzymology
chemical_list: D006846:Hydrocarbons, Halogenated; D009578:Nitrobenzenes; D009249:NADP; D009247:NADH, NADPH Oxidoreductases; D005978:Glutathione; D010100:Oxygen; D019803:Glutathione Disulfide