Abstract
The effects of acetone on liver, kidney, and lung monooxygenases were studied using hamsters administered 8% acetone in drinking water. Binding of aniline to liver microsomes induced a type II difference spectrum, and the spectral binding was enhanced in hamsters pretreated with acetone. Administration of acetone caused significant increases of cytochrome P-450 and cytochromeb 5 contents in liver microsomes. The increases of the hemeproteins were associated with induction of monooxygenase activities toward test substrates, aniline, N-nitrosodimethylamine, benzphetamine, benzo(a)pyrene, and 7-ethoxycoumarin. In the kidneys, acetone administration increased microsomal contents of the hemeprotein and monooxygenase activities toward aniline, N-nitrosodimethylamine, and 7-ethoxycoumarin, but not benzphetamine or benzo(a)pyrene. In the lungs, acetone pretreatment increased aniline hydroxylase activity without affecting the levels of N-nitrosodimethylamine demethylase, cytochromes P-450 andb 5. In marked contrast to the inductive effects in the liver, acetone administration markedly decreased lung microsomal benzo(a)pyrene hydroxylase and 7-ethoxycoumarin O-deethylase activities. Gel electrophoresis of liver and kidney microsomes from control and acetone-treated hamsters revealed that acetone treatment enhanced the intensity of a protein band(s) in the cytochrome P-450 molecular weight region. Immunoblotting of the microsomal proteins showed that the protein band induced by acetone in hamster liver, kidney and lung was cross-reactive with antibody raised against ethanol-inducible human liver cytochrome P-450. These results demonstrate that acetone has the ability to uniformly induce a specific form of cytochrome P-450, designated as IIE1, and to cause differential changes of monooxygenase activities in the hamster tissues. The complex effects of acetone on hepatic and extrahepatic monooxygenase systems may be important determinants of tissue-specific chemical toxicity.
Similar content being viewed by others
Abbreviations
- NDMA:
-
N-nitrosodimethylamine
- P-450:
-
cytochrome P-450
- bs :
-
cytochromebs
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
Ardies CM, Lasker JM, Lieber CS (1987) Characterization of the cytochrome P-450 monooxygenase system of hamster liver microsomes. Effects of prior treatment with ethanol and other xenobiotics. Biochem Pharmacol 36: 3613–3619
Bartsch H, Malaveille C, Montesano R (1975) In vitro metabolism and microsome-mediated mutagenicity of dialkylnitrosamines in rat, hamster, and mouse tissues. Cancer Res 35: 644–651
Bellward GD, Chang T, Rodrigues B, McNeill JH, Maines S, Ryan DE, Levin W, Thomas PE (1988) Hepatic cytochrome P-450j induction in the spontaneously diabetic BB rat. Mol Pharmacol 33: 140–143
Brady JF, Lee MJ, Li M, Ishizaki H, Yang S (1988) Diethyl ether as a substrate for acetone/ethanol-inducible cytochrome P-450 and as an inducer for cytochrome(s) P-450. Mol Pharmacol 33: 148–154
Casazza JP, Felver ME, Veech RL (1984) The metabolism of acetone in rat. J Biol Chem 259: 231–236
Conney AH (1982) Induction of microsomal enzymes by foreign chemicals and carcinogenesis by polycyclic aromatic hydrocarbons. Cancer Res 42: 4875–4917
Coon MJ, Koop DR (1987) Alcohol-inducible cytochrome P-450 (P-450ALC). Arch Toxicol 60: 16–21
Ding X, Koop DR, Crump BL, Coon MJ (1986) Immunochemical identification of cytochrome P-450 isozyme 3a (P-450ALC) in rabbit nasal and kidney microsomes and evidence for differential induction by alcohol. Mol Pharmacol 30: 370–378
Dong Z, Hong J, Ma Q, Li D, Bullock J, Gonzalez FJ, Park SS, Gelboin HV, Yang CS (1988) Mechanism of induction of cytochrome P-450ac (P-450j) in chemically induced and spontaneously diabetic rats. Arch Biochem Biophys 263: 29–35
Elves RG, Ueng T-H, Alvares AP (1984) Comparative effects of ethanol administration on hepatic monooxygenases in rats and mice. Arch Toxicol 55: 258–264
Fujii H, Ohmachi T, Sagami I, Wantabe M (1985) Liver microsomal drug metabolism in ethanol-treated hamsters. Biochem Pharmacol 34: 3881–3884
Greenlee WF and Poland A (1978) An improved assay of 7-ethoxycoumarin O-deethylase activity: induction of hepatic enzyme activity in C57BL/6J and DBA/2J mice by phenobarbital, 3-methylcholanthrene and 2,3,7,8-tetrachlorodibenzo-p-dioxin. J Pharmacol Exp Ther 205: 596–605
Guengerich FP (1989) Characterization of human microsomal cytochrome P-450 enzymes. Annu Rev Pharmacol Toxicol 29: 241–264
Hong J-Y, Pan J, Dong Z, Ning SM, Yang CS (1987a) Regulation of N-nitrosodimethylamine demethylase in rat liver and kidney. Cancer Res 47: 5948–5953
Hong J, Pan J, Gonzalez FJ, Gelboin HV, Yang CS (1987b) The induction of a specific form of cytochrome P-450 (P-450j) by fasting. Biochem Biophys Res Commun 142: 1077–1083
Imai Y, Ito A, Sato R (1966) Evidence for biochemically different types of vesicles in the hepatic fraction. J Biochem (Tokyo) 60: 417–428
Johansson I, Ekstrom G, Scholte B, Puzycki D, Jornvall H, Ingelman-Sundberg M (1988) Ethanol-, fasting-, and acetone-inducible cytochromes P-450 in rat liver: regulation and characteristics of enzymes belonging to the IIB and IIE gene subfamilies. Biochemistry 27: 1925–1934
Kimura S, Kozak CA, Gonzalez FJ (1989) Identification of a novel P450 expressed in rat lung: cDNA cloning and sequence, chromosome mapping, and induction by 3-methylcholanthrene. Biochemistry 28: 3798–3803
Koop DR, Casazza JP (1985) Identification of ethanol-inducible P-450 isozyme 3a as the acetone and acetol monooxygenase of rabbil microsomes. J Biol Chem 260: 13607–13612
Koop DR, Morgan ET, Tarr GE, Coon MJ (1982) Purification and characterization of a unique isozyme of cytochrome P-450 from liver microsomes of ethanol-treated rabbits. J Biol Chem 257: 8472–8480
Koop DR, Crump BL, Nordblom GD, Coon MJ (1985) Immunochemical evidence for induction of the alcohol-oxidizing cytochrome P-450 of rabbit liver microsomes by diverse agents: ethanol, imidazole, trichloroethylene, acetone, pyrazole, and isoniazid. Proc Natl Acad Sci USA 82: 4065–4069
Kubato S, Lasker JM, Lieber CS (1988) Molecular regulation of ethanolinducible cytochrome P450-IIE1 in hamsters. Biochem Biophys Res Commun 150: 304–310
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227: 680–685
Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein determination with Folin phenol reagent. J Biol Chem 193: 265–275
Lu AYH, West SB (1980) Multiplicity of mammalian microsomal cytochromes P-450. Pharmacol Rev 31: 277–295
Miller KW, Yang CS (1984) Studies on the mechanisms of induction of N-nitrosodimethylamine demethylase by fasting, acetone, and ethanol. Arch Biochem Biophys 229: 483–491
Nash T (1953) The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. Biochem J 55: 416–421
Nebert DW, Gelboin HV (1968) Substrate-inducible microsomal aryl hydroxylase in mammalian cell culture. I. Assembly and properties of induced enzyme. J Biol Chem 243: 6242–6249
Nebert DW, Adesnik M, Coon MJ, Estabrook RW, Gonzalez FJ, Guengerich FP, Gunalus IC, Johnson EF, Kemper B, Levin W, Phillips IR, Sato R, Waterman MR (1987) The P450 gene superfamily: recommended nomenclature. DNA 6: 1–11
Nebert DW, Nelson DR, Adesnik M, Coon MJ, Estabrook RW, Gonzalez FJ, Guengerich FP, Gunsalus IC, Johnson EF, Kemper B, Levin W, Phillips IR, Sato R, Waterman MR (1989) The P450 superfamily: updated listing of all genes and recommended nomenclature for the chromosomal loci. DNA 8: 1–13
Omura T, Sato R (1964) The carbon monoxide-binding pigment of liver microsomes. I. Evidence for its hemeprotein nature. J Biol Chem 239: 2370–2379
Patten CJ, Ning SM, Lu AYH, Yang CS (1986) Acetone-inducible cytochrome P-450: purification, catalytic activity, and interaction with cytochromeb 5. Arch Biochem Biophys 251: 629–638
Phillips AH, Langdon RG (1962) Hepatic triphosphopyridine nucleotidecytochromec reductase: isolation, characterization, and kinetic studies. J Biol Chem 237: 2652–2660
Plaa GL, Hewitt WR, Du Souich P, Caille G, Lock S (1982) Isopropanol and acetone potentiation of carbon tetrachloride-induced hepatotoxicity: single versus repetitive pretreatments in rats. J Toxicol Environ Health 9: 235–250
Porter TD, Khani SC, Coon MJ (1989) Induction and tissue-specific expression of rabbit cytochrome P450IIE1 and IIE2 genes. Mol Pharmacol 36: 61–65
Ryan DE, Thomas PE, Reik LM, Levin W (1982) Purification, characterization and regulation of five rat hepatic microsomal cytochrome P-450 isozymes. Xenobiotica 11: 727–744
Ryan DE, Ramanthan L, Iida S, Thomas PE, Haniu M, Shively JE, Lieber CS, Levin W (1985) Characterization of a major form of rat hepatic microsomal cytochrome P-450 induced by isoniazid. J Biol Chem 260: 6385–6393
Ryan DE, Koop DR, Thomas PE, Coon MJ, Levin W (1986) Evidence that isoniazid and ethanol induce the same microsomal cytochrome P-450 in rat liver, an isozyme homologous to rabbit liver cytochrome P-450 isozyme 3a. Arch Biochem Biophys 246: 633–644
Shimada T, Iwasaki M, Martin MV, Guengerich FP (1989) Human liver microsomal cytochrome P-450 enzymes involved in the bioactivation of procarcinogens detected byumu gene response inSalmonella typhimurium TA1535/pSK1002. Cancer Res 49: 3218–3228
Sipes IG, Stripp B, Krishna G, Maling HM, Gillette JR (1973) Enhanced hepatic microsomal activity by pretreatment of rats with acetone or isopropanol. Proc Soc Exp Biol Med 142: 237–240
Song B-J, Gelboin HV, Park S-S, Yang CS, Gonzalez FJ (1986) Complementary DNA and protein sequences of ethanol-inducible rat and human cytochrome P-450s. Transcriptional and post-transcriptional regulation of the rat enzyme. J Biol Chem 261: 16689–16697
Song B-J, Veech RL, Park SS, Gelboin HV, Gonzalez FJ (1989) Induction of rat hepatic N-nitrosodimethylamine demethylase by acetone is due to protein stabilization. J Biol Chem 264: 3568–3572
Thomas PE, Bandiera S, Maines SL, Ryan DE, Levin W (1987) Regulation of P-450j, a high affinity N-nitrosodimethylamine demethylase, in rat hepatic microsomes. Biochemistry 26: 2280–2289
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Tu YY, Peng R, Chang Z-F, Yang CS (1983) Induction of a high affinity nitrosamine demethylase in rat liver microsomes by acetone and isopropanol. Chem-Biol Interact 44: 247–260
Ueng T-H, Friedman FK, Miller H, Park SS, Gelboin HV, Alvares AP (1987) Studies on ethanol-inducible cytochrome P-450 in rabbit liver, lungs and kidneys. Biochem Pharmacol 6: 2689–2691
Vainio H, Hietanen E (1980) Role of extrahepatic metabolism. In: Jenner P, Testa B (eds) Concepts in drug metabolism, part A. Marcell Dekker Inc., New York, pp 251–284
Vanderslice RR, Domin BA, Carver GT, Philpot RM (1987) Species-dependent expression and induction of homologues of rabbit cytochrome P-450 isozyme 5 in liver and lung. Mol Pharmacol 31: 320–325
Widmark EMP (1920) Studies in the acetone concentration in blood, urine, and alveolar air. II: The passage of acetone and aceto-acetic acid into the urine. Biochem J 14: 364–378
Wrighton SA, Thomas PE, Molowa DT, Haniu M, Shively JE, Maines SL, Watkins PB, Parker G, Mendez-Picon G, Levin W, Guzelian PS (1986) Characterization of ethanol-inducible human liver N-nitrosodimethylamine demethylase. Biochemistry 25: 6731–6735
Wrighton SA, Thomas PE, Ryan DE, Levin W (1987) Purification and characterization of ethanol-inducible human hepatic cytochrome P-450HLj. Arch Biochem Biophys 258: 292–297
Yang CS, TU YY, Koop DR, Coon MJ (1985) Metabolism of nitrosamines by purified rabbit liver cytochrome P-450 isozymes. Cancer Res 45: 1140–1145
Yoo J-SH, Ning SM, Patten CJ, Yang CS (1987) Metabolism and activation of N-nitrosodimethylamine by hamster and rat microsomes: comparative study with weanling and adult animals. Cancer Res 47: 992–998
Author information
Authors and Affiliations
Additional information
The nomenclature of P-450 used in this report follows the system recommended by Nebert et al. (1987, 1989). P-450 IIE1 has also been referred to as P-450ac by Patten et al. (1986), P-450j by Ryan et al. (1986), and as isozyme 3a and P-450ALC by Coon and Koop (1987) in various species.
Rights and permissions
About this article
Cite this article
Ueng, TH., Tsai, JN., Ju, JM. et al. Effects of acetone administration on cytochrome P-450-dependent monooxygenases in hamster liver, kidney, and lung. Arch Toxicol 65, 45–51 (1991). https://doi.org/10.1007/BF01973502
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01973502