Target cells for cytochrome p450-catalysed irreversible binding of 7,12-dimethylbenz[a]anthracene (DMBA) in rodent adrenal glands.
7,12-Dimethylbenz[a]anthracene (DMBA) is an adrenocorticolytic agent that causes apoplexy (haemorrhage) and massive necrosis in the adrenal cortex in rat. Several explanations regarding the origin of toxicity have been proposed. Huggins and Morii (J Exp Med 114:741-60, 1961) suggested that the cells of the inner adrenal cortex are the primary target, whereas Horváth and Kovács (Pathol Eur 8:43-59, 1973) suggested the vascular endothelium as being the origin of toxicity. In the present study, cultured precision-cut tissue slices were used to localize target cells for irreversible [(3)H]DMBA binding in rat and mouse adrenal cortex. The sites of binding were confirmed by autoradiography in vivo. Irreversible [(3)H]DMBA binding was confined to zona fasciculata/reticularis cells in rat (but not in mouse) adrenal cortex. Pronounced binding was observed in clusters of cells (focal binding), localized predominantly in zona reticularis of rat. [(3)H]DMBA binding in zona fasciculata/reticularis cells was inhibited by the cytochrome p450 1A/B (CYP1A/B) inhibitors ellipticine, alpha-naphthoflavone, and 1-ethynylpyrene. The CYP11B1-inhibitor metyrapone did not reduce [(3)H]DMBA binding. In CYP1-induced (PCB 126-treated) rats and mice, intense irreversible [(3)H]DMBA binding was found also in endothelial cells of the adrenal cortex. The endothelial binding was abolished by the CYP1 inhibitors but remained unaffected by metyrapone. We conclude that the metabolic activation in adrenal parenchymal cells is presumably catalysed by CYP1B1, whereas CYP1A1 presumably catalyses the activation in endothelial cells. We suggest that the adrenocorticolytic effect of DMBA is the result of a dual mode of action, targeting both endothelial and parenchymal cells in the rat adrenal cortex.
Department of Environmental Toxicology, Evolutionary Biology Center, Uppsala University, Norbyvägen 18A, S-752 36 Uppsala, Sweden.
Orjan
O
Lindhe
Lizette
L
Granberg
Ingvar
I
Brandt
D015127:9,10-Dimethyl-1,2-benzanthracene / Q000378:metabolism*; D000302:Adrenal Cortex / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000037:antagonists & inhibitors; D001345:Autoradiography; D001571:Benzoflavones / Q000494:pharmacology; D001665:Binding Sites; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors; D065633:Cytochrome P-450 CYP1B1; D065607:Cytochrome P-450 Enzyme Inhibitors; D004611:Ellipticines / Q000494:pharmacology; D004727:Endothelium / Q000473:pathology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D051379:Mice; D008815:Mice, Inbred Strains; D011078:Polychlorinated Biphenyls / Q000494:pharmacology; D011721:Pyrenes / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015384:Zona Glomerulosa / Q000201:enzymology / Q000378:metabolism / Q000473:pathology; D015385:Zona Reticularis / Q000201:enzymology / Q000378:metabolism / Q000473:pathology
D001571:Benzoflavones; D065607:Cytochrome P-450 Enzyme Inhibitors; D004611:Ellipticines; D004791:Enzyme Inhibitors; D011721:Pyrenes; C034192:ellipticine; C042881:1-ethynylpyrene; D015127:9,10-Dimethyl-1,2-benzanthracene; C011512:alpha-naphthoflavone; D011078:Polychlorinated Biphenyls; D001189:Aryl Hydrocarbon Hydroxylases; C585498:Cyp1b1 protein, mouse; C585499:Cyp1b1 protein, rat; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1; C023035:3,4,5,3',4'-pentachlorobiphenyl