3,4-Dichlorophenylhydroxylamine cytotoxicity in renal cortical slices from Fischer 344 rats.
3,4-Dichlorophenylhydroxylamine (3,4-CPHA) is the N-hydroxyl metabolite of 3,4-dichloroaniline. 3,4-Dichloroaniline is a breakdown product of the herbicide Propanil. Previous work has shown that 3,4-dichloroaniline is acutely toxic to the kidney and bladder. The purpose of this study was to examine the in vitro toxicity of 3,4-dichlorophenylhydroxylamine. Renal cortical slices were prepared from male Fischer 344 rats (190-250 g) and were incubated with 0-0.5 mM 3,4-CPHA for 30-120 min under oxygen and constant shaking. 3,4-CPHA produced a concentration and time dependent alteration in lactate dehydrogenase (LDH) leakage, organic ion accumulation and pyruvate stimulated gluconeogenesis. Glutathione levels were diminished within 60 min below control values by 0.1 and 0.5 mM 3,4-CPHA. A 30 min pretreatment with 0.1 mM deferoxamine did not alter 3,4-CPHA toxicity. Alterations in pyruvate stimulated gluconeogenesis and LDH leakage were comparable between vehicle and deferoxamine pretreated tissues. Other studies examined the effect of (1 mM) glutathione, 2 mM ascorbic acid and 1 mM dithiothreitol (DTT) on toxicity. Pretreatment for 30 min with vehicle or 1 mM DTT induced comparable changes in LDH leakage and pyruvate stimulated gluconeogenesis. Pretreatment for 30 min with 1 mM glutathione or 2 mM ascorbic acid reduced 3,4-CPHA toxicity. LDH leakage was not elevated as markedly in renal slices pretreated with glutathione relative to slices pretreated with vehicle. These results indicate that 3,4-CPHA toxicity is through an iron independent mechanism. 3,4-CPHA cytotoxicity was reduced by pretreatment with glutathione or ascorbic acid suggesting formation of a reactive intermediate.
Valentovic
M
M
Department of Pharmacology, Marshall University School of Medicine, 1542 Spring Valley Drive, Huntington, WV 25704-9388, USA. valentov@marshall.edu
Ball
J G
JG
Stoll
S
S
Rankin
G O
GO
D000704:Analysis of Variance; D000814:Aniline Compounds / Q000494:pharmacology / Q000633:toxicity*; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005943:Gluconeogenesis / Q000187:drug effects; D005978:Glutathione / Q000494:pharmacology; D006898:Hydroxylamines / Q000494:pharmacology / Q000633:toxicity*; D066298:In Vitro Techniques; D007672:Kidney Cortex / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism / Q000473:pathology; D007674:Kidney Diseases / Q000139:chemically induced* / Q000201:enzymology* / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D019289:Pyruvic Acid / Q000378:metabolism; D051381:Rats; D011916:Rats, Inbred F344
D000814:Aniline Compounds; D006898:Hydroxylamines; C014464:3,4-dichloroaniline; C069709:3,4-dichlorophenylhydroxylamine; D019289:Pyruvic Acid; D007770:L-Lactate Dehydrogenase; D005978:Glutathione; D003676:Deferoxamine; D001205:Ascorbic Acid; D004229:Dithiothreitol