Alpha-oxidative metabolism of the bladder carcinogens N-nitrosobutyl(4-hydroxybutyl)amine and N-nitrosobutyl(3-carboxypropyl)amine within the rat isolated bladder.
The most widely accepted metabolic pathway leading to the formation of reactive intermediates from nitrosamines involves enzymatic hydroxylation at the carbon atom alpha to the nitroso moiety. All subsequent steps are non-enzymatic reactions and the final result is the stoichiometric formation of a cationic product and molecular nitrogen. Thus the amount of molecular nitrogen evolved can be used as an indicator of alpha-hydroxylation. The use of doubly 15N-labelled nitrosamines and the detection of 15N2 by MS makes it simpler to measure the extent of alpha-hydroxylation. We have studied the alpha-oxidation of doubly 15N-labelled N-nitrosobutyl(4-hydroxybutyl)amine (BBN) and its metabolite N-nitrosobutyl(3-carboxypropyl)amine (BCPN), two potent urinary bladder carcinogens in animals, within the target organ. Various amounts of 15N-labelled BBN ranging from 0.1 to 5 mumol were incubated at 37 degrees C for 4 h in the isolated rat bladder and the formation of 15N2 was measured by GC-MS. 15N2 production was linear up to 1 mumol and represented approximately 0.1% of the substrate incubated. Time-course experiments showed that 15N2 production was linear over a 6 h incubation period, ranging from 2.16 +/- 0.05 to 4.55 +/- 0.33 nmol/mg urothelial cell protein. 15N-labelled BCPN (1-5 mumol) was also incubated within the rat isolated bladder. 15N2 production from BCPN was approximately 10 times less than that from BBN. The results indicate that, though to a lower extent, the target organ activates 15N-labelled BBN and BCPN through the alpha-hydroxylation pathway.
Airoldi
L
L
Laboratory of Environmental Pharmacology and Toxicology, Istituto di Ricerche Farmacologiche Mario Negri, Milano, Italy.
Magagnotti
C
C
Bonfanti
M
M
Fanelli
R
R
D000818:Animals; D001711:Biotransformation; D002085:Butylhydroxybutylnitrosamine / Q000378:metabolism*; D002273:Carcinogens / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D009602:Nitrosamines / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D001743:Urinary Bladder / Q000378:metabolism*
D002273:Carcinogens; D009602:Nitrosamines; D002085:Butylhydroxybutylnitrosamine; C006853:butyl(3-carboxypropyl)nitrosamine