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In vitro modulation of the P450 activities of hamster and human lung slices

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Abstract

The respiratory tract is a portal of entry for many environmental chemicals. The respiratory tract plays an important role in the detoxification or metabolic activation of these chemicals, e.g., via cytochrome P450 enzymes. Alterations in the capabilities of these enzymes to metabolize inhaled compounds can, therefore, affect the toxicity of the chemicals. The pulmonary cytochrome P450 activity has been studied in many species, but relatively little is known about this activity in the human lung tissue. In this limited study, we have investigated the possibility of modulating in vitro the P450 activity in lung slices from hamsters and humans. The alkoxyresorufin-O-dealkylase activity was measured in the S9 fraction of lung slices incubated for 24 h with 106 mol/L 20-methylcholanthrene (3MC) or β-naphthoflavone (βN). The ethoxyresorufin-O-deethylase (EROD) activity was increased by 3MC and βN in lung slices of both species. The benzyloxyresorufin-O-deethylase (BROD) activity was decreased after incubation with 3MC but increased with βN. These data show that in vitro modulation in lung slices is feasible, although technical improvement is still needed, particularly in relation to the viability of the slices.

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References

  • Abdel-Razzak Z, Corcos L, Fautrel A, Guillouzo A. Interleu-kin-1 beta antagonizes phenobarbital induction of several major cytochromes P450 in adult rat hepatocytes in primary culture. FEBS Lett. 1995;366:159–64.

    Google Scholar 

  • Akrawi M, Rogiers V, Vandenberghe Y et al. Maintenance and induction in co-cultured rat hepatocytes of components of the cytochrome P450-mediated mono-oxygenase. Biochem Pharmacol. 1993;45:1583–91.

    Google Scholar 

  • Baron J, Voigt JM. Localization, distribution and induction of xenobiotic-metabolizing enzymes and aryl hydrocarbon hydroxylase activity within lung. In: Gram TE, ed. Meta-bolic activation and toxicity of chemical agents to lung tissue and cells. Oxford: Pergamon; 1994:41–76.

    Google Scholar 

  • Burke MD, Mayer RT. Ethoxyresorufin: direct fluorimetric assay of a microsomal O-dealkylation which is preferentially inducible by 3-methylcholanthrene. Drug Metab Dispos. 1974;2:583–8.

    Google Scholar 

  • Burke MD, Prough RA, Mayer RT. Characteristics of a microsomal cytochrome P-448-mediated reaction. Ethoxy-resorufin O-de-ethylation. Drug Metab Dispos. 1977;5:1–8.

    Google Scholar 

  • Burke MD, Thompson S, Elcombe CR, Halpert J, Haaparanta T, Mayer RT. Ethoxy-, pentoxy-and benzyloxyphenoxa-zones and homologues: a series of substrates to distinguish between different induced cytochromes P-450. Biochem Pharmacol. 1985;34:3337–45.

    Google Scholar 

  • Cardenas A, Nemery B. Effects of pneumotoxic trialkylphos-phorothioates on the pentose phosphate pathway in rat lung slices. Toxicol Lett. 1991;56:339–48.

    Google Scholar 

  • Charest M, Rossignol G, Castonguay A. In vitro and in vivo modulation of the bioactivation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in hamster lung tissues. Chem-Biol Interact. 1989;71:265–78.

    Google Scholar 

  • Curi-Pedrosa R, Daujat M, Pichard L et al. Omeprazole and lansoprazole are mixed inducers of CYP1A and CYP3A in human hepatocytes in primary culture. J Pharmacol Exp Ther. 1994;269:384–92.

    Google Scholar 

  • Devereux TR. Alveolar type II and Clara cells: isolation and xenobiotic metabolism. Environ Health Perspect. 1984;56: 95–101.

    Google Scholar 

  • Devereux TR, Fouts JR. Xenobiotic metabolism by alveolar type II cells isolated from rabbit lung. Biochem Pharmacol. 1981;30:1231–7.

    Google Scholar 

  • Devereux TR, Domin BA, Philpot RM. Xenobiotic metabolism by isolated pulmonary bronchiolar and alveolar cells. In: Gram TE, ed. Metabolic activation and toxicity of chemical agents to lung tissue and cells. Oxford: Pergamon; 1994:25–40.

  • Fisher AB, Reicherter J. Pentose pathway of glucose metabo-lism in isolated granular pneumocytes. Metabolic regulation and stimulation by paraquat. Biochem Pharmacol. 1984;33: 1349–53.

    Google Scholar 

  • Fisher RL, Smith MS, Hasal SJ, Hasal KS, Gandolfi AJ, Brendel K. The use of human lung slices in toxicology. Hum Exp Toxicol. 1994;13:466–71.

    Google Scholar 

  • Hoet PHM, Dinsdale D, Lewis CPL, Verbeken EK, Lauweryns JM, Nemery B. Kinetics and cellular localisation of putres-cine uptake in human lung. Thorax. 1993;48:1235–41.

    Google Scholar 

  • Howell BF, McCune S, Schaffer R. Lactate-to-pyruvate or pyruvate-to-lactate assay for lactate dehydrogenase: a re-examination. Clin Chem. 1979;25:269–72.

    Google Scholar 

  • Ishinishi N, Tanaka A, Hisanaga A, Inamasu T, Hirata M. Comparative study on the carcinogenicity of N-nitroso-diethylamine, N-nitrosodimethylamine, N-nitrosomorpho-line, N-nitrosopyrrolidine, N-nitrosodi-n-propylamine to the lung of Syrian golden hamsters following intermittent instillations to the trachea. Carcinogenesis. 1988;9:947–50.

    Google Scholar 

  • Lake BG, Beamand JA, Japenga AC, Renwick A, Davies S, Price RJ. Induction of cytochrome P-450-dependent en-zyme activities in cultured rat liver slices. Food Chem Toxic. 1993;31:377–86.

    Google Scholar 

  • Lake BG, Charzat C, Tredger JM, Renwick AB, Beamand JA, Price RJ. Induction of cytochrome P450 isoenzymes in cultured precision-cut rat and human liver slices. Xenobio-tica. 1996;26:297–306.

    Google Scholar 

  • Levy SE, Harvey E. Effect of tissue slicing on rat lung metabolism. J Appl Physiol. 1974;37:239–40.

    Google Scholar 

  • Lewis CPL, Demedts M, Nemery B. Indices of oxidative stress in hamster lung following exposure to cobalt(II) ions: in vivo and in vitro studies. Am J Respir Cell Mol Biol. 1991; 5:163–9.

    Google Scholar 

  • Lubet RA, Nims RW, Mayer RT, Cameron JW, Schechtman LM. Measurement of cytochrome P-450 dependent de-alkylation of alkoxy-phenoxazones in hepatic S9C and hepatocyte homogenates: effects of dicumarol. Mutat Res. 1985;142:127–31.

    Google Scholar 

  • Lubet RA, Syi JL, Nelson JO, Nims RW. Induction of hepatic cytochrome P-450 mediated alkoxyresorufin O-dealkylase activities in different species by prototype P-450 inducers. Chem-Biol Interact. 1990;75:325–39.

    Google Scholar 

  • McLemore TL, Martin RR, Busbee DL et al. Aryl hydrocarbon hydroxylase activity in pulmonary macrophages and lym-phocytes from lung cancer and noncancer patients. Cancer Res. 1977;37:1175–81.

    Google Scholar 

  • McLemore T, Martin RR, Pickard LR et al. Analysis of aryl hydrocarbon hydroxylase activity in human lung tissue, pulmonary macrophages, and blood lymphocytes. Cancer. 1978;41:2292–300.

    Google Scholar 

  • Miller MS, Warner SP, Jorquera R, Castonguay A, Schüller HM. Expression of the cytochrome P4502E and 2B gene families in the lungs and livers of nonpregnant, pregnant, and fetal hamsters. Biochem Pharmacol. 1992;44:797–803.

    Google Scholar 

  • Nebert DW, Nelson DR, Coon MJ et al. The P450 superfamily: update on new sequences, gene mapping, and recommended nomenclature. DNA Cell Biol. 1991a;10:1–14.

    Google Scholar 

  • Nebert DW, Petersen DD, Puga A. Human AH locus poly-morphism and cancer: inducibility of CYP1A1 and other genes by combustion products and dioxin. Pharmacoge-netics. 1991b;1:68–78.

    Google Scholar 

  • O'Neil JJ, Sanford RL, Wasserman S, Tierney DF. Metabolism in rat lung tissue slices: technical factors. J Appl Physiol. 1977;43:902–6.

    Google Scholar 

  • Overby LH, Nishio S, Weir A, Carver GT, Plopper CG, Philpot RM. Distribution of cytochrome P4501A1 and NADPH-cytochrome P450 reductase in lungs of rabbits treated with 2,3,7,8-tetrachlorodibenzo-p-dioxin: ultrastructural immu-nolocalization and in situ hybridization. Mol Pharmacol. 1992;41:1039-4

    Google Scholar 

  • Rabovsky J, Judy DJ. Cytochrome P450-dependent alkoxyphenoxazone dealkylase activities in lung microsomes fromuntreated and β-naphthoflavone-treated rats; effect of in vitro inhibitors. Res Commun Chem Pathol Pharmacol.1987;57:375-87.

    Google Scholar 

  • Sagami I, Ohmachi T, Fujii H, Kikuchi H, Watanabe M. Hamster cytochrome P-450IA gene family, P-450IA1 and P-450IA2 in lung and liver: cDNA cloning and sequence analysis. J Biochem (Tokyo). 1991;110:641–7.

    Google Scholar 

  • Strum JM, Ito T, Philpot RM, DeSanti AM, McDowell EM. The immunocytochemical detection of cytochrome P-450 monooxygenase in the lungs of fetal, neonatal, and adult hamsters. Am J Respir Cell Mol Biol. 1990;2:493–501.

    Google Scholar 

  • Vassault A. Lactate dehydrogenase. 1. UV-method with pyr-uvate and NADH}. In: Methods of enzymatic analyses. Vol III: Enzymes I: oxidireductase, transferase, 3rd ed. Wein-heim: Verlag Chemie; 1983:118–26.

    Google Scholar 

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Authors and Affiliations

  1. Laboratorium voor Pneumologie, K. U. Leuven, Leuven, Belgium

    P.H.M. Hoet, M. Demedts & B. Nemery

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  1. P.H.M. Hoet
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  2. M. Demedts
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  3. B. Nemery
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Hoet, P., Demedts, M. & Nemery, B. In vitro modulation of the P450 activities of hamster and human lung slices. Cell Biol Toxicol 13, 185–192 (1997). https://doi.org/10.1023/A:1007362329736

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