Metabolism Of 1-nitropyrene Research Articles

Metabolism of 1-nitropyrene in mice: transport across the placenta and mammary tissues

The distribution and metabolism of the environmental pollutant 1-nitropyrene was studied in C57B1/6N mice following oral or intraperitoneal dosing. When administered by gavage, 1-nitropyrene and its metabolites demonstrated biphasic elimination kinetics from the blood, with half-lives of 0.3 and 1.8 days and a distribution volume of 74 ml. Intraperitoneal administration resulted in similar biphasic elimination, with half-lives of 0.5 and 3 days and a distribution volume of 98 ml. Treating pregnant C57B1/6N (C 2H sire) mice by gavage resulted in similar absorption and elimination kinetics of 1-nitropyrene and metabolites, except that the distribution volume increased to 123 ml. 1-Nitropyrene and/or its metabolites (0.7% of the administered dose) crossed the placenta and accumulated in the fetuses and amniotic fluid, with both C-oxidized and nitroreduced metabolites being detected. Suckling neonates accumulated 1-nitropyrene and its metabolites when their dams were administered 1-nitropyrene by gavage. Each neonate received approximately 0.1% of the administered dose and demonstrated the presence of both C-oxidized and nitroreduced metabolites. These results demonstrate that this environmental pollutant is capable of crossing the placenta or mammary tissues to expose the offspring to a potentially genotoxic compound.

Effect of enzyme inducers on metabolism of 1-nitropyrene in human hepatoma cell line HepG2

We measured the response of HepG2 cells to the classic cytochrome (cyt.) P-450 inducers 3-methylcholanthrene (3-MC) and phenobarbital (PB), by evaluating oxidative and/or reductive metabolism of the nitroarenes, 1-NP and 1,6-dinitropyrene (1,6-DNP), in control and induced cells. In HepG2 cells, 3-MC induces ring-hydroxylation of 1-NP, whereas PB stimulates its nitroreduction. PB induces NADPH-cyt. c reductase, but does not affect other cytosolic and microsomal enzymes which contribute to 1-NP nitroreduction in these cells. However, PB-inducible nitroreductase activity seems to be associated primarily with cyt. P-450 isoenzymatic form(s), as indicated by the requirement for NADPH and the response to specific inhibitors such as α-naphthoflavone and CO.

The metabolism of 1-nitropyrene by human cytochromes P450

The metabolism of [3H]1-nitropyrene by specific forms of human cytochrome P450 was investigated in vitro using Vaccinia virus expression of P450 cDNAs in HepG2 cells. Cell lysates were injected individually with recombinant Vaccinia virus containing human P450 cDNA (P450 IA2, IIA3, IIB7, IIC8, IIC9, IID6, IIE1, IIF1, IIIA3, IIIA4, IIIA5 AND IVB1). Only IIIA3 and IIIA4 demonstrated significant activity in the C-oxidation of 1-nitropyrene. The principal metabolite from both P450 forms was 1-nitropyren-3-ol, produced in at least 4-fold greater abundance than the mixture of 1-nitropyren-6-ol and 1-nitropyren-8-ol, or the K-region dihydrodiols. This is in contrast to the metabolism in many species where 6-ol and 8-ol formation predominate over 3-ol formation. In fact, in rats and rabbits, P450 forms quite distinct from the IIIA P450s catalyze the majority of the metabolism of this pollutant. This is the first demonstration of the role of individual human P450 forms in the metabolism of a representative chemical in this important class of environmental pollutants. The importance of these observations in the overall carcinogenic risk of humans to these chemicals remains to be established. These studies furthermore establish a marked species difference in the metabolism of nitrated polycyclic aromatic hydrocarbons.

In vitro intestinal microflora-mediated metabolism of biliary metabolites from 1-nitropyrene-treated rats.

To investigate the modifying role of the intestinal microflora in the metabolism of 1-nitropyrene (1-NP) via enterohepatic circulation, we collected bile from male Wistar rats administered [3H]1-NP orally. The bile was mixed with the intestinal contents (IC) prepared from untreated rats and the mixture was incubated anaerobically under an atmosphere of nitrogen at 37 C. Samples of the reaction mixture were removed at intervals to assay their mutagenic potential, to determine the radioactivity bound to the IC, and for analysis of the biliary metabolites. The binding of the radioactivity to the IC increased linearly as a function of time during the 1-hr incubation. The time-dependent binding does not occur with heat-treated IC and the binding was inhibited by addition of D-saccharic acid 1,4-lacton, a beta-glucuronidase inhibitor. The mutagenicity (for Salmonella typhimurium strain TA98 without S9 mix) of the bile increased early in the incubation period and then decreased very rapidly. The mutagenicity of the bile was also enhanced by treatment with a sonicated IC extract or beta-glucuronidase, but not with a heat-treated IC or aryl-sulfatase. The metabolites produced after the bile was incubated for short periods with the IC were mainly nitrohydroxypyrenes; at later times nitroreduction occurred. The level of acetylaminohydroxypyrenes, which were formed by deconjugation, did not change during the incubation. To determine the degree of contribution of the IC to the total acetylating capacity, we measured acetyltransferase activity of the IC and various organs in Wistar rats. The liver had the highest N-acetyltransferase activity among the seventeen organs examined. Considerable activity was also detected in the kidney, small intestine, lung, and testis, but the IC showed very low activity. The acetylating capacity of the IC was 0.27% of the total capacity in rats, and that of the liver was more than 80%. These results suggest that the nitrohydroxypyrenes formed from 1-NP in the liver were conjugated to glucuronic acid and excreted via the bile duct into intestine. Hydrolysis of these glucuronide conjugates by bacterial beta-glucuronidase liberated into intestine, free nitrohydroxypyrenes, which were direct-acting mutagens. The released aglycons were then rapidly nitro-reduced by intestinal microflora, but contribution of the intestinal microflora to acetylation of the reduced metabolites is very low.

In vivo metabolism of 1-nitropyrene: detection of glutathione conjugates of 1-nitropyrene 4,5-oxide in the bile of rats administered 1-nitropyrene orally

In vivo metabolism of 1-nitropyrene: detection of glutathione conjugates of 1-nitropyrene 4,5-oxide in the bile of rats administered 1-nitropyrene orally

Metabolism of 1-nitropyrene by cultured rabbit alveolar macrophages and respiratory tract tissues

The metabolism of 1-nitro[ 14C]pyrene ( 14C-1-NP; 8.1 μ m) was studied in cultured (20 hr) rabbit alveolar macrophages, lung tissue, and tracheal tissue. Metabolites from the incubation medium and from the macrophages and respiratory tract tissues were extracted and then analyzed and quantified by high-pressure liquid chromatography. The following metabolites were detected in the lung and tracheal tissue incubation medium: 1-nitropyrene-4,5-dihydrodiol, N-acetyl-1-aminopyrene, 1-aminopyrene, and 10-hydroxy-1-nitropyrene. Nitropyrene phenols (4-, 5-, 6-, 8- or 9-hydroxy-1-nitropyrene) and 3-hydroxy-1-nitropyrene were only detected in the lung and tracheal tissue and not in the incubation medium for these tissues. Minor amounts of 1-aminopyrene and 10-hydroxy-1-nitropyrene were detected in the macrophage incubation medium, and only minute quantities of 1-nitropyrene-4,5-dihydrodiol, 1-aminopyrene, and 10-hydroxy-1-nitropyrene were detected in macrophages. The total percentage of 1-NP metabolism was significantly greater in the lung and tracheal tissue (28.0 and 23.0% of the recovered 14C, respectively) than in the alveolar macrophages (6.3% of the recovered 14C). The tracheal tissue was found to have the highest activity both in 1-NP metabolism and intracellular metabolite concentration. A major portion of the 1-NP metabolites produced was released into the incubation medium. The majority of the metabolites produced by tracheal and lung tissue, 70 and 84%, respectively, were ethyl acetate extractable. The metabolites retained within the cells or tissues were also predominantly ethyl acetate extractable rather than water soluble (83% for the macrophages and trachea, 95% for the lung tissue). The metabolite profiles obtained demonstrate that metabolism by both nitro reduction and ring oxidation occurs in respiratory tissue, and a degree of tissue specificity in the formation of metabolites exists. Ring oxidation was demonstrated in the lung and tracheal tissue, but very little occurred in the macrophages.

Metabolism of 1-nitropyrene in germfree and conventional rats

Metabolism of 1-nitropyrene in germfree and conventional rats

Excretion and metabolism of 1-nitropyrene in rats after oral or intraperitoneal administration

Many nitro-substituted polycyclic aromatic hydrocarbons (NPAHs) have been identified as environmental pollutants and have been found to be mutagens and carcinogens in bacteria and mammalian systems. They require metabolism to express their biological activity. The metabolism and excretion of 1-nitropyrene (NP), a prevalent NPAH, by Fischer-344 rats after intraperitoneal (ip) or oral administration was studied. Radiolabeled NP was administered to rats (10 mg NP/kg body wt), and urine and feces were collected for 7 days. After ip administration of [ 14C]NP, 60% of the radioactivity was found in the urine and 20% in the feces. Likewise, 55 and 35% of the orally administered 14C was found in urine and feces, respectively. Both urine and feces were analyzed by high-pressure liquid chromatography for metabolites. The majority of the radioactivity in both urine and feces was associated with very polar metabolites, none accounting for more than 10% of the dose. Small amounts (less than 1% of the dose) of aminopyrene (AP), acetylaminopyrene, and NP were detected. A urinary metabolite (3–8% of the dose) was found that converted to acetylaminopyrene phenol (two isomers) when urine was heated overnight at 37°C at pH 4.5. More of this metabolite (2.2 times) as well as AP (1.8 times), was excreted after oral than after ip administration of NP. The NP metabolites found in this study demonstrate that reduction of the nitro group is a significant route of NP metabolism in rats. Since nitroreduction appears to be necessary in the activation of NPAHs to bacterial mutagens, this indicates that similar metabolic pathways are present in rats (catalyzed by mammalian and/or gut bacterial enzymes) and that activation of NPAHs to carcinogens or toxins by nitroreduction is possible.

Excretion and Metabolism of 1-Nitropyrene in Rats after Oral or Intraperitoneal Administration

Excretion and Metabolism of 1-Nitropyrene in Rats after Oral or Intraperitoneal Administration

Tumorigenicity and metabolism of 1-nitropyrene in A/J mice.

Four groups of 28-32 male and female A/J mice were given i.p. injections of either trioctanoin or 1-nitropyrene in trioctanoin such that the total doses of 1-nitropyrene were 0.71 mmol/kg, 2.14 mmol/kg, or 6.44 mmol/kg. The mean number of lung tumors/mouse was 1.3 +/- 1.0 in the group treated with 6.44 mmol/kg of 1-nitropyrene compared with 0.3 +/- 0.6 in the trioctanoin group (p less than 0.001). Combined tumor incidence was not significant compared with controls in the two lower dose groups. Cultured explants of A/J mouse lung, and 9000 g supernatant of A/J mouse lung and liver, metabolized [14C]1-nitropyrene to 4,5-dihydro-4,5-dihydroxy-1-nitropyrene and 1-nitrohydroxypyrenes. Substantial amounts of unknown polar metabolites were also produced by cultured lung. Nitro-reduction to 1-aminopyrene was minimal in mouse lung and liver, even under oxygen deficient conditions.

Metabolism of 1-nitropyrene and formation of DNA adducts in Salmonella typhimurium.

1-Nitropyrene is slowly reduced by intact cells of Salmonella typhimurium to yield 1-aminopyrene and N-acetyl-1-aminopyrene plus six unidentified minor products. When the bacteria are exposed to tritiated 1-nitropyrene, increasing amounts of radioactivity become bound to DNA as the nitropyrene is metabolized. Enzymatic hydrolysis of the labelled DNA yields low molecular weight labelled compounds which probably represent nucleoside adducts formed by the reaction of nitropyrene metabolites with DNA. Results with appropriate mutant strains indicate that bacterial nitroreductases are involved in activating nitropyrene to a reactive intermediate that binds to DNA and that nitropyrene adducts in DNA are subject to excision repair.