Cytochrome P450-dependent binding of 7,12-dimethylbenz[a]anthracene (DMBA) and benzo[a]pyrene (B[a]P) in murine heart, lung, and liver endothelial cells.
Autoradiography was used to investigate the cellular sites of irreversible binding of 3H-labelled 7,12-dimethylbenz[a]anthracene (DMBA) and benzo[a]pyrene (B[a]P) in mice. Autoradiograms obtained from solvent-extracted tape-sections revealed an even distribution of DMBA- and B[a]P-derived radioactivity in control mice lacking sites of selective binding in the tissues. In mice pretreated with a cytochrome P4501A (CYP1A) inducer, beta-naphthoflavone (BNF) or 3,3',4,4', 5-pentachlorobiphenyl (PCB 126), a noticeable accumulation of bound radioactivity was observed in the pulmonary alveolar region. Increased labelling was also observed in heart tissue of induced mice. As demonstrated by microautoradiography of tissues from CYP1A-induced mice treated with 3H-DMBA or 3H-B[a]P in vivo, irreversible binding in lung tissue was present in endothelial cells of arteries and veins, in the alveolar septal walls, and in type 2 pneumocytes. In heart tissue, binding was confined to endothelial cells of arteries, capillaries and veins. In liver, binding was found in the hepatocytes as well as in endothelial cells of the portal veins, whereas no binding was seen in endothelial cells of the sinusoids, central veins, or arteries. These findings were confirmed in vitro using 3H-DMBA-exposed precision-cut slices, indicating that reactive intermediates of DMBA and B(a)P were formed in situ. The addition of the CYP1A inhibitor ellipticine abolished binding in the target endothelial cells. Increased endothelial binding in the lungs and liver of CYP1A-induced mice was concomitant with increased 7-ethoxyresorufin O-deethylase (EROD) and DMBA hydroxylase activity. In heart, endothelial binding was positively correlated with EROD, but not with DMBA hydroxylase. The results suggest that endothelial cells may be targets for CYP-dependent activation of such toxicants as polycyclic aromatic hydrocarbons. Consequently, the possibility that chemically induced endothelial dysfunction is a risk factor in the aetiology of cardiovascular disease demands consideration.
Granberg
A L
AL
Department of Environmental Toxicology, Evolutionary Biology Centre, Uppsala University, Sweden.
Brunström
B
B
Brandt
I
I
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000096:biosynthesis*; D001345:Autoradiography; D001551:Benz(a)Anthracenes / Q000493:pharmacokinetics*; D001564:Benzo(a)pyrene / Q000493:pharmacokinetics*; D002273:Carcinogens / Q000493:pharmacokinetics*; D003331:Coronary Vessels / Q000166:cytology / Q000378:metabolism; D019363:Cytochrome P-450 CYP1A1 / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D004611:Ellipticines / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D004785:Environmental Pollutants / Q000493:pharmacokinetics*; D004790:Enzyme Induction; D005260:Female; D066298:In Vitro Techniques; D008099:Liver / Q000098:blood supply; D008168:Lung / Q000098:blood supply; D051379:Mice; D011078:Polychlorinated Biphenyls / Q000494:pharmacology; D014018:Tissue Distribution; D019324:beta-Naphthoflavone / Q000494:pharmacology
D001551:Benz(a)Anthracenes; D002273:Carcinogens; D004611:Ellipticines; D004785:Environmental Pollutants; C034192:ellipticine; C046810:7,12-dihydroxymethylbenz(a)anthracene; D001564:Benzo(a)pyrene; D019324:beta-Naphthoflavone; D011078:Polychlorinated Biphenyls; C037450:7,12-dimethylbenz(a)anthracene hydroxylase; D001189:Aryl Hydrocarbon Hydroxylases; D019363:Cytochrome P-450 CYP1A1; C023035:3,4,5,3',4'-pentachlorobiphenyl