Lymphocyte cytochrome P450 expression: inducibility studies in male Wistar rats.

Mary P.A Hannon-Fletcher, Yvonne A. Barnett

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The cytochrome P450 system plays a key role in the metabolism of endogenous and exogenous compounds. The system is widely distributed in body tissues with the highest concentration of the enzymes found in the liver hepatocyte. Extra hepatic expression of the P450 system has been documented in the lung, pancreas and kidney and the enzymes have been shown to be induced by many disease states, including diabetes mellitus and cancer. Little attention has been paid to the expression and inducibility of the system in peripheral blood lymphocytes. In this present investigation, specific P450 inducers were administered in vivo to male Wistar rats. The expression and in vivo induction of the P450 isoforms; CYP2B, CYP2E, CYP3A and CYP4A, within liver and lymphocyte samples was determined using Western blot analysis. Following in vivo induction the lymphocyte P450 proteins showed an average 3-fold increase in expression (0.003-0.005 g P450/g microsomal protein), compared to the control lymphocyte samples. Expression in the induced lymphocyte samples was up to 11-fold lower than that of the induced liver samples, as expected. Results indicate that lymphocytes may provide a relatively simple method for monitoring the P450 profile in human subjects.
LanguageEnglish
Pages1-5
JournalBritish Journal of Biomedical Science
Volume65
Issue number1
Publication statusPublished - 2008

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Cytochrome P-450 Enzyme System
Wistar Rats
Lymphocytes
Liver
Cytochrome P-450 CYP4A
Cytochrome P-450 CYP3A
Enzymes
Pancreas
Hepatocytes
Diabetes Mellitus
Protein Isoforms
Proteins
Western Blotting
Kidney
Lung
Neoplasms

Cite this

@article{9443dd497e014e7bab716457db070137,
title = "Lymphocyte cytochrome P450 expression: inducibility studies in male Wistar rats.",
abstract = "The cytochrome P450 system plays a key role in the metabolism of endogenous and exogenous compounds. The system is widely distributed in body tissues with the highest concentration of the enzymes found in the liver hepatocyte. Extra hepatic expression of the P450 system has been documented in the lung, pancreas and kidney and the enzymes have been shown to be induced by many disease states, including diabetes mellitus and cancer. Little attention has been paid to the expression and inducibility of the system in peripheral blood lymphocytes. In this present investigation, specific P450 inducers were administered in vivo to male Wistar rats. The expression and in vivo induction of the P450 isoforms; CYP2B, CYP2E, CYP3A and CYP4A, within liver and lymphocyte samples was determined using Western blot analysis. Following in vivo induction the lymphocyte P450 proteins showed an average 3-fold increase in expression (0.003-0.005 g P450/g microsomal protein), compared to the control lymphocyte samples. Expression in the induced lymphocyte samples was up to 11-fold lower than that of the induced liver samples, as expected. Results indicate that lymphocytes may provide a relatively simple method for monitoring the P450 profile in human subjects.",
author = "Hannon-Fletcher, {Mary P.A} and Barnett, {Yvonne A.}",
note = "Reference text: 1. Yang CS. and Lu AYH. The diversity of substrates for cytochrome P-450. In: Guengerich. FP, ed. Mammalian Cytochrome P-450, Volume 2. Boca Raton , USA: CRC Press, 1987:1-18. 2. Nelson DR, Koymans L, Stegeman JJ, Feyereisen R, Waxman DJ, Waterman MR, Gotoh O, Coon MJ, Estabrook RW, Gunsalus IC, Nebert DW. P450 superfamily: update on new sequences, gene mapping, accession numbers and nomenclature. Pharmacogenetics 1996; 6: 1-42. 3. Gonzalez FJ, Gelboin FJ. Role of human cytochrome P-450 in the metabolic activation of chemical carcinogens and toxins. Drug Met Rev 1994; 26: 165-183. 4. Gonzalez FJ. The molecular biology of cytochrome P450s. Pharm. Rev 1989; 40: 243-288. 5. Nebert DW. Role of Genetics and Drug Metabolism in Human Cancer Risk. Mutat Res 1991; 247: 267-281 6. Ioannides C, Barnett CR, Flatt PR. The effects of experimental diabetes on the cytochrome P450 system and other metabolic pathways. In: McNeill JH, ed. Experimental Models of Diabetes. Boca Raton, USA: CRC Press, 1999: 79-116 7. Barnett CR, Flatt PR, Ioannides C. Hyperketonaemia markedly modulates the metabolic activation of chemical carcinogens. Chemo-Biol Inte 1990a; 74: 281-289. 8. Lewis DF, Ioannides C, Parke DV. Cytochrome P450 and species differences in xenobiotic metabolism and activation of carcinogen. Environ Health Pers 1998; 106: 633-641. 9. Ennever FK, Noonan TJ, Rosenkranz HS. The predictivity of animal bioassays and short -term genotoxicity tests for carcinogenicity and non-carcinogenicity to humans. Mutagenesis 1987; 2: 73-78. 10. Lijinsky W. Life-span and cancer: the induction time of tumours in diverse animal species treated with nitrosodiethylamine. Carcinogenesis 1993; 14: 2373-2375. 11. Barnett CR, Flatt PR, Ioannides C. Induction of hepatic microsomal P450 I and IIB proteins by hyperketonaemia. Biochem Pharma 1990b; 40: 393-397. 12. Barnett CR, Gibson GG, Wolf CR, Flatt PR, Ioannides C. Induction of cytochrome P450III and P450IV family proteins in streptozotocin-induced diabetes. Biochem J 1990c; 286: 765-769. 13. Ioannides C, Bass SL, Ayrton AD, Trinick J, Walker R, Flatt PR. Streptozotocin-induced diabetes modulates the metabolic activation of chemical carcinogens. Chemico-Biolo Inter 1988; 68: 189-202. 14. Irizar A, Ioannides C. Extrahepatic expression of P450 proteins in insulin-dependent diabetes mellitus. Xenobiotica 1995; 25: 941-949. 15. Clarke J, Flatt PR, Barnett CR. Cytochrome P450 CYP2E expression in the pancreatic islets of Langerhans. Diabetic Med 1996; 13: (Suppl. 3) 37. 16. Clarke J, Flatt PR, Barnett CR. Cytochrome P450 1A-like proteins expressed in the islets of Langerhans and altered pancreatic -cell secretory responsiveness. B J Pharma 1997; 121: 389-394. 17. Roberts BJ, Shoaf SE, Jeong KS, Song BJ. Induction of CYP2E in liver, kidney, brain and intestine during chronic ethanol administration and withdrawal: evidence that CYP2E possesses a rapid phase half-life of 6 hours or less. Biochem Biophys Res Comm 1994; 205: 1064-1071. 18. Kivisto KT, Fritz P, Linder A, Friedel G, Beaune P, Kroemer HK. Immunohistochemical localization of cytochrome P450 3A in human pulmonary carcinomas and normal bronchial tissue. Histochem 1995; 103: 25-29 19. Barnett CR, Flatt PR, Ioannides C. Role of ketone bodies in the diabetes-induced changes in hepatic mixed-function oxidase activities. Biochem et Bioph Acta 1988; 967: 250-254. 20. Barnett CR, Flatt PR, Bone AJ. Hepatic cytochrome P450 profile in BB rats with spontaneous insulin-dependent diabetes mellitus. In: Lechner MC, ed. Biochemistry, Biophysics, and Molecular Biology of Cytochrome P450. Paris: John Libbey Eurotext, 1994: 574-550. 21. Ioannides C, Barnett CR, Irizar A, Flatt PR. Drug metabolism and chemical toxicity in diabetes In: Ioannides C, Flatt PR, eds. Drugs, Diet and Disease Volume 2. Hemel Hempstead, UK: Ellis Horwood, 1995: 330-353. 22. Mahnke A, Roos PH, Hanstein WG, Chabot GG. In vivo induction of cytochrome P450 CYP3A expression in rat leukocytes using various inducers. Biochem Pharma 1996; 51:1579-1582. 23. Hannon-Fletcher MP, O’Kane M J, Moles KW, Barnett CR, Barnett YA. Lymphocyte CYP2E1 expression in human IDDM subjects. Food Chem Tox 2001; 39: 25-32 24. Song BJ, Veech RL, Saenger P. Cytochrome P450IIE1 is elevated in lymphocytes from poorly controlled insulin-dependant diabetics. J. Clin Endoc Metab 1990; 71: 1036-1040. 25. Raucy JL, Schultz ED, Wester MR, Arora S, Johnston DE, Omdahl JL, Carpenter SP. Human lymphocyte cytochrome P450 2E1, a putative marker for alcohol-mediated changes in hepatic chlorzoxazone activity. Drug Meta Dispos 1997; 25: 1429-1435. 26. Kouri RE, McKinney CE, Slomiany DJ, Snodgrass DR, Wray NP, McLemore TL. Positive correlation between high aryl hydrocarbon hydroxylase activity and primary lung cancer as analysed in cryopreserved lymphocytes. Cancer Res 1982; 42: 5030-5037. 27. Gurtoo HL, Parker NB, Paigen B, Havens MB, Minowada J, Freeman HJ. Induction, inhibition, and some enzymological properties of aryl hydrocarbon hydroxylase in fresh mitogen-activated human lymphocytes. Cancer Res 1979; 39: 4620-4629. 28. Kellermann G, Luyten-Kellermann M, Shaw CR. Genetic variation of aryl hydrocarbon hydroxylase in human lymphocytes. Amer J Hum Genet 1973; 25: 237-331. 29. Rumsby PC, Yardley-Jones A, Anderson D, Phillimore HE, Davies MJ. Detection of CYP1A1 mRNA levels and CYP1A1 Msp polymorphisms as possible biomarkers of exposure and susceptibility in smokers and non-smokers. Teratog Carcinog Mutagen 1996; 16: 65-74. 30. Lee P-C, Werlin SL. The induction of hepatic cytochrome P450 3A in rats: effects of age. Pro Soc Exper Biol Med 1995; 210: 134-139. 31. Ioannides C, Parke DV. Mechanism of induction of hepatic microsomal drug metabolising enzymes by a series of barbiturates. J Pharmac Pharma 1975; 27: 739-746. 32. B{\"o}yum A. Isolation of mononuclear cells and granulocytes from human blood. Norwegian Defence Research Establishment, Division for Toxicology. (Paper iv) Kjeller, Norway, 1980: 77-89. 33. Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principal protein-dye binding. Anal Biochem 1976; 72: 248-252. 34. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685. 35. Merk HF, Baron J, Hertl M, Niederau D, R{\"u}bben A. Lymphocyte activation in allergic reactions elicited by small-molecular-weight compounds. Inter Arch Allerg Immun 1997; 113: 173-176. 36. Murray GI, Barnes TS, Sewell HF, Ewen SW, Melvin WT, Burke MD. The immunocytochemical localisation and distribution of cytochrome P450 in normal human hepatic and extra hepatic tissues with a monoclonal antibody to human cytochrome P450. B J Clin Pharma 1988; 25: 465-475. 37. Bondy SC, Naderi S. Contribution of hepatic cytochrome P450 systems to the generation of reactive oxygen species. Biochem Pharma 1994; 48: 155-159. 38. Oberley LW, Oberley TD. Reactive oxygen species in the aetiology of cancer. In: Ioannides C, Lewis DFV, eds. Drugs, Diet and Disease. Volume 1. Mechanistic approaches to cancer. London: Ellis Horwood, 1995: 47-63. 39. Baynes JW. Reactive oxygen in the aetiology and complications of diabetes. In: Ioannides C, Flatt PR, eds. Drugs, Diet & Disease Volume 2. Mechanistic Approaches to Diabetes. Hemel Hempstead: Ellis Hoilwood, 1995: 201-240.",
year = "2008",
language = "English",
volume = "65",
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}

Lymphocyte cytochrome P450 expression: inducibility studies in male Wistar rats. / Hannon-Fletcher, Mary P.A; Barnett, Yvonne A.

In: British Journal of Biomedical Science, Vol. 65, No. 1, 2008, p. 1-5.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Lymphocyte cytochrome P450 expression: inducibility studies in male Wistar rats.

AU - Hannon-Fletcher, Mary P.A

AU - Barnett, Yvonne A.

N1 - Reference text: 1. Yang CS. and Lu AYH. The diversity of substrates for cytochrome P-450. In: Guengerich. FP, ed. Mammalian Cytochrome P-450, Volume 2. Boca Raton , USA: CRC Press, 1987:1-18. 2. Nelson DR, Koymans L, Stegeman JJ, Feyereisen R, Waxman DJ, Waterman MR, Gotoh O, Coon MJ, Estabrook RW, Gunsalus IC, Nebert DW. P450 superfamily: update on new sequences, gene mapping, accession numbers and nomenclature. Pharmacogenetics 1996; 6: 1-42. 3. Gonzalez FJ, Gelboin FJ. Role of human cytochrome P-450 in the metabolic activation of chemical carcinogens and toxins. Drug Met Rev 1994; 26: 165-183. 4. Gonzalez FJ. The molecular biology of cytochrome P450s. Pharm. Rev 1989; 40: 243-288. 5. Nebert DW. Role of Genetics and Drug Metabolism in Human Cancer Risk. Mutat Res 1991; 247: 267-281 6. Ioannides C, Barnett CR, Flatt PR. The effects of experimental diabetes on the cytochrome P450 system and other metabolic pathways. In: McNeill JH, ed. Experimental Models of Diabetes. Boca Raton, USA: CRC Press, 1999: 79-116 7. Barnett CR, Flatt PR, Ioannides C. Hyperketonaemia markedly modulates the metabolic activation of chemical carcinogens. Chemo-Biol Inte 1990a; 74: 281-289. 8. Lewis DF, Ioannides C, Parke DV. Cytochrome P450 and species differences in xenobiotic metabolism and activation of carcinogen. Environ Health Pers 1998; 106: 633-641. 9. Ennever FK, Noonan TJ, Rosenkranz HS. The predictivity of animal bioassays and short -term genotoxicity tests for carcinogenicity and non-carcinogenicity to humans. Mutagenesis 1987; 2: 73-78. 10. Lijinsky W. Life-span and cancer: the induction time of tumours in diverse animal species treated with nitrosodiethylamine. Carcinogenesis 1993; 14: 2373-2375. 11. Barnett CR, Flatt PR, Ioannides C. Induction of hepatic microsomal P450 I and IIB proteins by hyperketonaemia. Biochem Pharma 1990b; 40: 393-397. 12. Barnett CR, Gibson GG, Wolf CR, Flatt PR, Ioannides C. Induction of cytochrome P450III and P450IV family proteins in streptozotocin-induced diabetes. Biochem J 1990c; 286: 765-769. 13. Ioannides C, Bass SL, Ayrton AD, Trinick J, Walker R, Flatt PR. Streptozotocin-induced diabetes modulates the metabolic activation of chemical carcinogens. Chemico-Biolo Inter 1988; 68: 189-202. 14. Irizar A, Ioannides C. Extrahepatic expression of P450 proteins in insulin-dependent diabetes mellitus. Xenobiotica 1995; 25: 941-949. 15. Clarke J, Flatt PR, Barnett CR. Cytochrome P450 CYP2E expression in the pancreatic islets of Langerhans. Diabetic Med 1996; 13: (Suppl. 3) 37. 16. Clarke J, Flatt PR, Barnett CR. Cytochrome P450 1A-like proteins expressed in the islets of Langerhans and altered pancreatic -cell secretory responsiveness. B J Pharma 1997; 121: 389-394. 17. Roberts BJ, Shoaf SE, Jeong KS, Song BJ. Induction of CYP2E in liver, kidney, brain and intestine during chronic ethanol administration and withdrawal: evidence that CYP2E possesses a rapid phase half-life of 6 hours or less. Biochem Biophys Res Comm 1994; 205: 1064-1071. 18. Kivisto KT, Fritz P, Linder A, Friedel G, Beaune P, Kroemer HK. Immunohistochemical localization of cytochrome P450 3A in human pulmonary carcinomas and normal bronchial tissue. Histochem 1995; 103: 25-29 19. Barnett CR, Flatt PR, Ioannides C. Role of ketone bodies in the diabetes-induced changes in hepatic mixed-function oxidase activities. Biochem et Bioph Acta 1988; 967: 250-254. 20. Barnett CR, Flatt PR, Bone AJ. Hepatic cytochrome P450 profile in BB rats with spontaneous insulin-dependent diabetes mellitus. In: Lechner MC, ed. Biochemistry, Biophysics, and Molecular Biology of Cytochrome P450. Paris: John Libbey Eurotext, 1994: 574-550. 21. Ioannides C, Barnett CR, Irizar A, Flatt PR. Drug metabolism and chemical toxicity in diabetes In: Ioannides C, Flatt PR, eds. Drugs, Diet and Disease Volume 2. Hemel Hempstead, UK: Ellis Horwood, 1995: 330-353. 22. Mahnke A, Roos PH, Hanstein WG, Chabot GG. In vivo induction of cytochrome P450 CYP3A expression in rat leukocytes using various inducers. Biochem Pharma 1996; 51:1579-1582. 23. Hannon-Fletcher MP, O’Kane M J, Moles KW, Barnett CR, Barnett YA. Lymphocyte CYP2E1 expression in human IDDM subjects. Food Chem Tox 2001; 39: 25-32 24. Song BJ, Veech RL, Saenger P. Cytochrome P450IIE1 is elevated in lymphocytes from poorly controlled insulin-dependant diabetics. J. Clin Endoc Metab 1990; 71: 1036-1040. 25. Raucy JL, Schultz ED, Wester MR, Arora S, Johnston DE, Omdahl JL, Carpenter SP. Human lymphocyte cytochrome P450 2E1, a putative marker for alcohol-mediated changes in hepatic chlorzoxazone activity. Drug Meta Dispos 1997; 25: 1429-1435. 26. Kouri RE, McKinney CE, Slomiany DJ, Snodgrass DR, Wray NP, McLemore TL. Positive correlation between high aryl hydrocarbon hydroxylase activity and primary lung cancer as analysed in cryopreserved lymphocytes. Cancer Res 1982; 42: 5030-5037. 27. Gurtoo HL, Parker NB, Paigen B, Havens MB, Minowada J, Freeman HJ. Induction, inhibition, and some enzymological properties of aryl hydrocarbon hydroxylase in fresh mitogen-activated human lymphocytes. Cancer Res 1979; 39: 4620-4629. 28. Kellermann G, Luyten-Kellermann M, Shaw CR. Genetic variation of aryl hydrocarbon hydroxylase in human lymphocytes. Amer J Hum Genet 1973; 25: 237-331. 29. Rumsby PC, Yardley-Jones A, Anderson D, Phillimore HE, Davies MJ. Detection of CYP1A1 mRNA levels and CYP1A1 Msp polymorphisms as possible biomarkers of exposure and susceptibility in smokers and non-smokers. Teratog Carcinog Mutagen 1996; 16: 65-74. 30. Lee P-C, Werlin SL. The induction of hepatic cytochrome P450 3A in rats: effects of age. Pro Soc Exper Biol Med 1995; 210: 134-139. 31. Ioannides C, Parke DV. Mechanism of induction of hepatic microsomal drug metabolising enzymes by a series of barbiturates. J Pharmac Pharma 1975; 27: 739-746. 32. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Norwegian Defence Research Establishment, Division for Toxicology. (Paper iv) Kjeller, Norway, 1980: 77-89. 33. Bradford MM. A rapid and sensitive method for the quantification of microgram quantities of protein utilising the principal protein-dye binding. Anal Biochem 1976; 72: 248-252. 34. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685. 35. Merk HF, Baron J, Hertl M, Niederau D, Rübben A. Lymphocyte activation in allergic reactions elicited by small-molecular-weight compounds. Inter Arch Allerg Immun 1997; 113: 173-176. 36. Murray GI, Barnes TS, Sewell HF, Ewen SW, Melvin WT, Burke MD. The immunocytochemical localisation and distribution of cytochrome P450 in normal human hepatic and extra hepatic tissues with a monoclonal antibody to human cytochrome P450. B J Clin Pharma 1988; 25: 465-475. 37. Bondy SC, Naderi S. Contribution of hepatic cytochrome P450 systems to the generation of reactive oxygen species. Biochem Pharma 1994; 48: 155-159. 38. Oberley LW, Oberley TD. Reactive oxygen species in the aetiology of cancer. In: Ioannides C, Lewis DFV, eds. Drugs, Diet and Disease. Volume 1. Mechanistic approaches to cancer. London: Ellis Horwood, 1995: 47-63. 39. Baynes JW. Reactive oxygen in the aetiology and complications of diabetes. In: Ioannides C, Flatt PR, eds. Drugs, Diet & Disease Volume 2. Mechanistic Approaches to Diabetes. Hemel Hempstead: Ellis Hoilwood, 1995: 201-240.

PY - 2008

Y1 - 2008

N2 - The cytochrome P450 system plays a key role in the metabolism of endogenous and exogenous compounds. The system is widely distributed in body tissues with the highest concentration of the enzymes found in the liver hepatocyte. Extra hepatic expression of the P450 system has been documented in the lung, pancreas and kidney and the enzymes have been shown to be induced by many disease states, including diabetes mellitus and cancer. Little attention has been paid to the expression and inducibility of the system in peripheral blood lymphocytes. In this present investigation, specific P450 inducers were administered in vivo to male Wistar rats. The expression and in vivo induction of the P450 isoforms; CYP2B, CYP2E, CYP3A and CYP4A, within liver and lymphocyte samples was determined using Western blot analysis. Following in vivo induction the lymphocyte P450 proteins showed an average 3-fold increase in expression (0.003-0.005 g P450/g microsomal protein), compared to the control lymphocyte samples. Expression in the induced lymphocyte samples was up to 11-fold lower than that of the induced liver samples, as expected. Results indicate that lymphocytes may provide a relatively simple method for monitoring the P450 profile in human subjects.

AB - The cytochrome P450 system plays a key role in the metabolism of endogenous and exogenous compounds. The system is widely distributed in body tissues with the highest concentration of the enzymes found in the liver hepatocyte. Extra hepatic expression of the P450 system has been documented in the lung, pancreas and kidney and the enzymes have been shown to be induced by many disease states, including diabetes mellitus and cancer. Little attention has been paid to the expression and inducibility of the system in peripheral blood lymphocytes. In this present investigation, specific P450 inducers were administered in vivo to male Wistar rats. The expression and in vivo induction of the P450 isoforms; CYP2B, CYP2E, CYP3A and CYP4A, within liver and lymphocyte samples was determined using Western blot analysis. Following in vivo induction the lymphocyte P450 proteins showed an average 3-fold increase in expression (0.003-0.005 g P450/g microsomal protein), compared to the control lymphocyte samples. Expression in the induced lymphocyte samples was up to 11-fold lower than that of the induced liver samples, as expected. Results indicate that lymphocytes may provide a relatively simple method for monitoring the P450 profile in human subjects.

M3 - Article

VL - 65

SP - 1

EP - 5

JO - British Journal of Biomedical Science

T2 - British Journal of Biomedical Science

JF - British Journal of Biomedical Science

SN - 0967-4845

IS - 1

ER -