Summary
Acute effects of a single intraperitoneal dose of allyl alcohol (AA, 64 mg/kg), dimethylnitrosamine (DMNA, 30 mg/kg), hexachlorobutadiene (HCBD, 50 mg/kg), carbon tetrachloride (CCl4, 24 mg/kg), cocaine (60 mg/kg) and pyrazole (300 mg/kg) on the hepatic histology and monooxygenases in DBA/2 and C57B1/6 strains of mice were investigated. All substances caused histologically verified injury to the liver, which varied in appearance and severity depending on the compound and the mouse strain. Responses of P450-catalyzed reactions were highly dependent on the toxin and varied between different monooxygenase (MO) reactions and two mouse strains. In DBA/2 strain, coumarin 7-hydroxylase (COH) activity was increased from 3- to 5-fold by pyrazole, cocaine, HCBD and CCl4. With respect to P450 content and other MO activities, no changes or even decreases were generally observed. Some exceptions to this rule were found: HCBD significantly increased T15αOH, PROD and EROD activities in C57B1/6 mice, whereas cocaine caused a significant stimulation of Tl5αOH and PROD in DBA/2 mice, It is concluded that (i) different hepatoxins cause different types of liver injury and responses of the monooxygenase complex (“hepatotoxin-specific finger prints”), (ii) although DBA/2 and C57B1/6 mice responded rather similarly to hepatotoxins, also with respect to P450 content and most MO activities, they displayed a profound difference in the behaviour of COH activity, and (iii) within the P450 superfamily, the regulation of COH activity seems to be rather unique, also when compared to its structurally close enzyme, testosterone 15α-hydroxylase.
Similar content being viewed by others
Abbreviations
- COH:
-
coumarin 7-hydroxylase
- P450Coh:
-
a P450 isozyme active in coumarin 7-hydroxylation
- T15 α OH:
-
testosterone 15α-hydroxylase
- P450 T 15 α OH:
-
a P450 enzymes catalyzing testosterone 15α-hydroxylation
- EROD:
-
7-ethoxyresorufin O-deethylase
- PROD:
-
7-pentoxyresorufin O-dealkylase
- AHH:
-
aryl hydrocarbon hydroxylase
- MO:
-
monooxygenase
- SDS:
-
sodium dodecyl sulphate
- DMNA:
-
dimethylnitrosamine
- CCl4 :
-
carbon tetrachloride
- HCBD:
-
hexachloro-1,3-butadiene
- AA:
-
allyl alcohol
References
Aida K, Negishi M (1991) Posttranscriptional regulation of coumarin 7-hydroxylase induction by xenobiotics in mouse liver: mRNA stabilization by pyrazole. Biochemistry 30:8041–8045
Aitio AA (1978) Simple and sensitive assay of 7-hydroxycoumarin deethylation. Anal Biochem 85:488–491
Arvela P, Kraul H, Stenbäck F, Pelkonen O (1991) The cerium-induced liver injury and oxidative drug metabolism in DBA/2 and C57BL/6 mice. Toxicology 69:1–9
Baron J, Voigt JM; Whitter TB, Kawabata TT, Knapp SA, Guengerich P, Jakoby WB (1986) Identification of intratissue sites for xenobiotic activation and detoxication. In: Nelson JO, Snyder R (eds) Biological reactive intermediates III. Plenum, New York
Bengtsson G, Julkunen A, Penttilä KE, Lindros KO (1987) Effect of Phenobarbital on the distribution of drug metabolizing enzymes between periportal and perivenous rat hepatocytes prepared by digitonin-collagenase liver perfusion. J Pharmacol Exp Ther 240: 663–667
Brambilla G, Carlo P, Finollo R (1992) Effect of ten thiocompounds on rat liver DNA damage induced by a small dose of N-nitrosodimethylamine. Arch Toxicol 66:286–290
Burke MD, Prough RA, Mayer RT (1977) Characteristics of a microsomal cytochrome P-448-mediated reaction ethoxyresorufin O-deethylation. Drug Metab Disp 5:1–8
Cohen AJ (1979) Critical review of the toxicology of coumarin with special reference to interspecies difference in metabolism and hepatotoxic response and their significance to man. Food Cosmet Toxicol 17:277–289
Egan D, O'Kennedy R, Moran E, Cox D, Prosser E, Thornes RD (1990) The pharmacology, metabolism, analysis, and applications of coumarin and coumarin-related compounds. Drug Metab Rev 22:503–529
Gascon-Barre M, Benbrahim N, Tremblay C (1989) Hepatic zonation of metabolizing enzymes. Studies on hepatocytes isolated from the periportal or perivenous region of the liver acinus. Can J Physiol Pharmacol 67:1015–1022
Gebhardt R (1992) Metabolic zonation of the liver: regulation and implications for liver function. Pharmacol Ther 53:275–354
Gonzalez FJ (1988) The molecular biology of cytochrome P450s. Pharmacol Rev 40:243–288
Greus Z, Watkins JB, Thompson TN, Klaassen CD (1982) Resistance of some phase II biotransformation pathways to hepatotoxins. J Pharm Exp Ther 222:471–479
Hahnemann B, Kuhn B, Heubel F, Legrum W (1989) Selective induction of the coumarin hydroxylase by N-containing heteroaromatic compounds. Arch Toxicol [Suppl 13]:297–301
Hahnemann B, Salonpää P, Pasanen M, Mäenpää J, Honkakoski P, Juvonen R, Lang MA, Pelkonen O, Raunio H (1992) Effect of pyrazole, cobalt and phenobarbital on Cyp 2a-4/5 (Cytochrome P450 2a-4/5) expression in the liver of C57BL/6 and DBA/2 mice. Biochemical J 286:289–294
Head B, Moody DE, Woo CH, Smuckler EA (1981) Alterations of specific forms of cytochrome P-450 in rat liver during acute tetrachloride intoxication. Tox Appl Pharmacol 61:286–295
Honkakoski P, Auriola S, Lang MA (1992) Distinct induction profiles of three phenobarbital-responsive mouse liver cytochrome P450 isozymes, Biochem Pharmacol 43:2121–2128
Honkakoski P, Lang MA (1989) Mouse liver phenobarbital-inducible P450 system: Purification, characterization and differential inducibility of four cytochrome P450 isozymes from the D2 mouse. Arch Biochem Biophys 273:42–57
James R, Desmond P, Kupfer A, Schenker S, Branch RA (1981) The differential localization of various drug metabolizing systems within the rat liver lobule as determined by the hepatotoxins allyl alcohol, carbon tetrachloride and bromobenzene. J Pharmacol Exp Ther 217:127–132
Juvonen RO, Autio S, Lang MA (1987) Pyrazole as a modifier of liver microsomal monooxygenase in BDA/2N and AKR/J mice. Biochem Pharmacol 36:3993–3997
Juvonen RO, Kaipainen PK, Lang MA (1985) Selective induction of coumarin 7-hydroxylase by pyrazole in D2 mice. Eur J Biochem 152:3–8
Kanai K, Kanamura S, Watanabe J (1986) Peri- and postnatal development of heterogeneity in the amounts of endoplasmic reticulum in mouse hepatocytes. Am J Anat 175:471–480
Kanai K, Watanabe J, Kanamura S (1990) Microphotometric analysis of cytochrome P450 in periportal, midzonal and perivenular hepatocytes of mice treated with phenobarbital. J Histochem Cytochem 38:1599–1605
Kloss MW, Rosen G, Rauchman EJ (1982) Acute cocaine-induced hepatotoxicity in DBA/2Ha male mice. Toxicol Appl Pharmacol 65:75–83
Kocer Z, Raunio H, Pasanen M, Arvela P, Raiskila T, Honkakoski P, Lang MA, Negishi M, Pelkonen O (1991) Comparison between cobalt and pyrazole in the increased expression of coumarin 7-hydroxylase in mouse liver. Biochem Pharmacol 41:462–465
Kojo A, Heiskanen R, Rytkönen A-L, Honkakoski P, Juvonen R, Lang M (1991) Inducibility of P450Coh by pyrazole and its derivatives. Biochem Pharmacol 42:1751–1759
Laemmli UK (1970) Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227:680–685
Lang MA, Juvonen RO, Järvinen P, Honkakoski P, Raunio H (1989) Mouse liver P450Coh: Genetic regulation of the pyrazole-inducible enzyme and comparison with other P450 isozymes. Arch Biochem Biophys 271:139–148
Lange R, Perin F, Larroque C, Zajdela F (1990) Isolation and partial characterization of a cytochrome P-450 isozyme (cytochrome P-450tu) from mouse liver tumors. Biochim Biophys Acta 1038: 130–135
Legrum W, Netter KJ (1980) Characteristics of coumarin metabolism by liver microsomes from cobalt-pretreated mice. Xenobiotica 10: 271–279
Letteron P, Labbe G, Degott C, Berson A, Fromenty B, Delaforge M, Larrey D, Pessayre D (1990) Mechanism for the protective effects of silymarin against carbon tetrachloride-induced lipid peroxidation and hepatotoxicity in mice. Evidence that silymarin acts both as an inhibitor of metabolic activation and as achain-braking antioxidant. Biochem Pharmacol 39:2027–2034
Lindberg R, Burkhart B, Ichikawa T, Negishi M (1989) The structure and characterization of type I P-45015α gene as major steroid 15α-hydroxylase and its comparison with type II P-45015α gene. J Biol Chem 264:6465–6471
Lush IE, Arnold CJ (1975) High coumarin 7-hydroxylase does not protect mice against warfarin, Heredity 35:279–281
Mangoura SA, Strack A, Legrum W, Netter KJ (1989) Indium selectively increases the cytochrome P-450 dependent O-dealkylation of coumarin derivatives in male mice. Naunyn-Schmiedeberg's Arch Pharmacol 339:596–602
Marshall WJ, McLean AEM (1969) The effect of chirrosis of the liver on microsomal detoxications and cytochrome P-450. Br J Exp Pathol 50:578–583
Moody DE, Taylor LA, Smuckler EA (1986) Immunohistochemical evidence for alteration in specific forms of rat hepatic microsomal cytochrome P-450 during acute carbon tetrachloride intoxication. Drug Metab Disp 14:709–713
Nebert DW, Gelboin HV (1968) Substrate-inducible microsomal aryl hydroxylase in mammalian cell culture. I. Assay and properties of induced enzymes. J Biol Chem 243:6242–6249
Nebert DW, Negishi M, Lang MA, Hjelmeland LM, Eisen JH (1982) The Ah locus, a multiple family necessary for survival in a chemically adverse environment: comparison with the immune system. Adv Genet 21:1–52
Nebert DW Robinson JR, Niwa A, Kumaki K, Poland AP (1975) Genetic expression of aryl hydrocarbon hydroxylase activity in the mouse. J Cell Physiol 85:393–414
Negishi M, Lindberg R, Burkhart B, Ichikawa T, Honkakoski P, Lang MA (1989) Mouse steroid 15α-hydroxylase gene family: Identification of type II P-45015, as a coumarin 7-hydroxylase. Biochemistry 28:4169–4172
Nogushi T, Fong KL, Lai EK, Olson L, McCcay PB (1982) Selective early loss of polypeptides in liver microsomes of CC14-treated rats. Biochem Pharmacol 31:609–614
Okino T, Nakajima T, Nakano M (1991) Morphological and biochemical analyses of trichloroethylene hepatotoxicity: Differences in ethanol-and phenobarbital-pretreated rats. Toxicol Appl Pharmacol 108:379–389
Omura T, Sato R (1964) The carbon monoxide-binding pigment of liver microsomes. I. Evidence for its hemoprotein nature. J Biol Chem 239:2370–2378
Raunio H, Kojo A, Juvonen R, Honkakoski P, Järvinen P, Lang MA, Vähäkangas K, Gelboin HV, Park SS, Pelkonen O (1988) Mouse hepatic cytochrome P-450 isozyme induction by 1,4- bis 2-(3,5-dichloropyridyloxy) benzene, pyrazole, and phenobarbital. Biochem Pharmacol 37:4141–4147
Salonpää P, Iscan M, Pasanen M, Arvela P, Pelkonen O, Raunio H (1992) Cerium-induced strain-dependent increase in Cyp2a-4/5 (Cytochrome P4502a-4/5) expression in the liver and kidneys of inbred mice. Biochem Pharmacol 44:1269–1274
Seibert B, Oesch F, Steinberg P (1989) Distribution and induction of cytochrome P450 and two cytochrome P450-dependent monooxygenase activities in rat liver parenchymal cell subpopulations separated by centrifugal elutriation. Arch Toxicol 63:18–22
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC (1985) Measurement of protein using bicinchronic acid. Anal Biochem 150:76–85
Smuckler EA, Arrhenius E, Hultin T (1967) Alterations in microsomal electron transport, oxidative N-demethylation and Azo-dye cleavage in carbon tetrachloride and dimethylnitrosamine-induced liver injury. Biochem J 103:55–64
Sohn DH, Yun YP, Park KS, Veech RL, Song BJ (1991) Post-translational reduction of cytochrome P450IIE by CC14, its substrate. Biochem Biophys Res Commun 179:449–454
Squires EJ, Negishi M (1988) Reciprocal regulation of sex-dependent expression of testosterone 15O-hydroxylase (P-45015a) in liver and kidney of male mice by androgen, evidence for a single gene. J Biol Chem 263:4166–4171
Thorgeirsson SS, Mitchell JR, Sasame HA, Potter WZ (1976) Biochemical changes after hepatic injury by allyl alcohol and N-hydroxy-2-acetylaminofluorene. Chem-Biol Interact 15:139–147
Towbin H, Staehulin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Väänanen H (1986) The distribution of cytochrome P450 mediated drug oxidation and glutathione in periportal and perivenous rat hepatocytes after phenobarbital treatment. J Hepatol 2:174–181
Waxman DJ, Ko A, Walsh C (1983) Regioselectivity and stereoselectivity of androgen hydroxylations catalyzed by cytochrome P-450 isozymes purified from phenobarbital-induced rat liver. J Biol Chem 258:11937–11947
Wood AW (1979) Genetic regulation of coumarin hydroxylase activity in mice. Biochemical characterization of the enzyme from two inbred strains and their F1 hybrid. J Biol Chem 254:5641–5646
Wood AW, Conney AH (1974) Genetic variation in coumarin hydroxylase activity in the mouse (Mus musculus). Science 185:612–614
Yamano S, Tatsuno J, Gonzalez FJ (1990) The CYP2A3 gene product catalyses coumarin 7-hydroxylation in human liver microsomes. Biochemistry 29:1322–1329
Author information
Authors and Affiliations
Additional information
An update list and suggested nomenclature of the sequenced P450 genes has been published recently (Nelson et al., DNA Cell Biol 12:1–51, 1993). The suggested nomenclature is followed in this manuscript. The murine P450Coh is the product of the Cyp2a-5 gene (the human counterpart is CYP2A6) and the murine P450T 15 α OH is the product of the Cyp2a-4 gene
Correspondence to: M. Pasanen at the above address
Rights and permissions
About this article
Cite this article
Pellinen, P., Stenbäck, F., Rautio, A. et al. Response of mouse liver coumarin 7-hydroxylase activity to hepatotoxins: dependence on strain and agent and comparison to other monooxygenases. Naunyn-Schmiedeberg's Arch Pharmacol 348, 435–443 (1993). https://doi.org/10.1007/BF00171345
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00171345