Abstract
Drug biotransformation and its therapeutic effect may be modified during ageing. Among different causative factors of ageing, the impairment of normal cellular functions by free radicals has been evoked as playing a critical role. The effect of age on the expression and activity of CYP2E1 and CYP3A was investigated in male Wistar rats of 3, 8, 11 and 18 months old. The total cytochrome P450 as well as the expression and the activity (midazolam oxidation) of CYP3A isoforms did not change until 18 months of age. Chlorzoxazone hydroxylation (CYP2E1 activity) increased from 3 to 8 months, remained constant between 8 and 11 months and then progressively decreased until 18 months. Interestingly, CYP2E1 microsomal protein followed the same enzyme activity profile from 3 to 8 months, but remained constant thereafter. The level of CYP2E1 mRNA did not change over the whole period. While the amount of proteins did not change after 8 months, their functionality may be affected by oxidative stress (increase in thiobarbituric acid reactive substances, decrease in reduced glutathione level). However, no changes in carbonyl protein content were observed. The decrease in CYP2E1 activity in rats after 11 months is most probably due to post-translational modifications of CYP2E1 proteins. Indeed, it may be correlated with an accumulation of oxidative damage. Since no change was observed in CYP3A activity or in their protein and mRNA content, it seems that such isoforms should be less affected by oxidative stress.
Similar content being viewed by others
References
Balducci L (2000) Geriatric oncology: challenges for the new century. Eur J Cancer 36:1741–1754
Birnbaum LS (1980) Altered hepatic drug metabolism in senescent mice. Exp Gerontol 15:259–267
Bogdanov M, Brown RH, Matson W, Smart R, Hayden D, O’Donnell H, Flint Beal M, Cudkowicz M (2000) Increased oxidative damage to DNA in ALS patients. Free Radic Biol Med 29:652–658
Bulteau A-L, Petropoulos I, Friguet B (2000) Age-related alterations of proteasomes structure and function in aging epidermis. Exp Gerontol 35:767–777
Bulteau A-L, Szweda LI, Friguet B (2002) Age-dependant declines in proteasome activity in the heart. Arch Biochem Biophys 397:298–304
Dodds C (1995) Anaesthetic drugs in the elderly. Pharmacol Ther 66:369–386
Dukan S, Farewell A, Ballesteros M, Taddei F, Radman M, Nystrom T (2000) Protein oxidation in response to increased transcriptional or translational errors. Proc Natl Acad Sci USA 97:5746–5749
Eeckhoudt SL, Desager JP, Horsmans Y, De Winne AJ, Verbeeck RK (1998) Sensitive assay for midazolam and its metabolite 1′-hydroxymidazolam in human plasma by capillary high-performance liquid chromatography. J Chromatogr B 710:165–171
Ekström G, Ingelman-Sundberg M (1989) Rat liver microsomal NADPH-supported oxidase activity and lipid peroxidation dependent on ethanol-inducible cytochrome P-450 (P-450IIE1). Biochem Pharmacol 38:1313–1319
Eliasson E, Mkrtchian S, Ingelman-Sunberg M (1992) Hormone- and substrate-regulated intracellular degradation of cytochrome P450 (2E1) involving MgATP-activated rapid proteolysis in the endoplasmic reticulum membranes. J Biol Chem 267:15765–15769
Espínola EB, Oliveira MGM, Carlini EA (1999) Differences in central and peripheral responses to oxotremorine in young and aged rats. Pharmacol Biochem Behav 62:419–423
Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408:239–247
Friguet B, Szweda LI, Stadtman ER (1994) Susceptibility of glucose-6-phosphate dehydrogenase modified by 4-hydroxy-2-nonenal and metal catalyzed oxidation to proteolysis by multicatalytic protease. Arch Biochem Biophys 311:168–173
George J, Byth K, Farrell GC (1995) Age but not gender selectively affects expression of individual cytochrome P450 proteins in human liver. Biochem Pharmacol 50:727–730
Goasduff T, Cederbaum AI (1999) NADPH-dependent microsomal electron transfer increases degradation of CYP2E1 by the proteasome complex: role of reactive oxygen species. Arch Biochem Biophys 370:258–270
Goasduff T, Cederbaum AI (2000) CYP2E1 degradation by in vitro reconstituted systems: role of the molecular chaperone hsp90. Arch Biochem Biophys 379:321–330
Grune T, Shringarpure R, Sitte N, Davies K (2001) Age-related changes in protein oxidation and proteolysis in mammalian cells. J Gerontol 56:459–467
Guengerich FP (1999) Cytochrome P450 3A4: regulation and role in drug metabolism. Annu Rev Pharmacol Toxicol 39:1–17
Hissin PJ, Hilf R (1976) A fluorometric method for determination of oxidized and reduced glutathione in tissues. Anal Biochem 74:214–226
Hunt CM, Strater S, Stave GM (1990) Effect of normal aging on the activity of human hepatic cytochrome P450IIE1. Biochem Pharmacol 40:1666–1669
Imaoka S, Fujita S, Funae Y (1991) Age-dependent expression of cytochrome P-450s in rat liver. Biochim Biophys Acta 1097:187–192
Kamataki T, Maeda K, Shimada M, Kitani K, Nagai T, Kato R (1985) Age-related alteration in the activities of drug-metabolizing enzymes and contents of sex-specific forms of cytochrome P-450 in liver microsomes from male and female rats. J Pharmacol Exp Ther 233:222–228
Kobayashi K, Urashima K, Shimada N, Chiba K (2002) Substrate specificity for rat cytochrome P450 (CYP) isoforms: screening with cDNA-expressed systems of the rat. Biochem Pharmacol 63:889–896
Leclercq I, Saliez A, Wallemacq PE, Horsmans Y, Lambotte L (1997) The monoethylglycinexylide test does not correctly evaluate lidocaine metabolism after ischemic liver injury in the rat. Hepatology 26:1182–1188
Leclercq I, Horsmans Y, Desager JP (1998) Estimation of chlorzoxazone hydroxylase activity in liver microsomes and of the plasma pharmacokinetics of chlorzoxazone by the same high-performance liquid chromatographic method. J Chromatogr A 828:291–296
Levine RL, Stadtman ER (2001) Oxidative modification of proteins during aging. Exp Gerontol 36:1495–1502
Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, Ahn BW, Shaltiel S, Stadtman ER (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478
Lowry OH, Rosebrough NJ, Farr AL, Randall R (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Mahnke A, Strotkamp D, Roos PH, Hanstein WG, Chabot GG, Nef P (1997) Expression and inducibility of cytochrome P450 3A9 (CYP3A9) and other members of the CYP3A subfamily in rat liver. Arch Biochem Biophys 337:62–68
Nakajima T, Wang RS, Katakura Y, Kishi R, Elovaara E, Park SS, Gelboin H, Vainio H (1992) Sex-, age- and pregnancy-induced changes in the metabolism of toluene and trichloroethylene in rat liver in relation to the regulation of cytochrome P450IIE1 and P450IIC11 content. J Pharmacol Exp Ther 261:869–887
Oesch-Bartlomowicz B, Oesch F (2003) Cytochrome-P450 phosphorylation as a functional switch. Arch Biochem Biophys 409:228–234
Omura T, Sato R (1964) The carbon monoxide-binding pigment of liver microsomes. II Solubilization, purification and properties. J Biol Chem 239:2370–2378
Rikans LE (1989) Hepatic drug metabolism in female Fischer rats as a function of age. Drug Metab Dispos 17:114–116
Rikans LE, Notley BA (1982) Age-related changes in hepatic microsomal drug metabolism are substrate selective. J Pharmacol Exp Ther 220:574–578
Ronis MJJ, Lindros KO, Ingelman-Sundberg M (1996) The CYP2E subfamily. In: Ioannides C (ed) Cytochromes P450: metabolic and toxicological aspects. CRC Press, Boca Raton, pp 211–239
Schmucker DL (2001) Liver function and phase I drug metabolism in the elderly: a paradox. Drugs Aging 18:837–851
Schmucker DL, Wang RK (1981) Effects of aging and phenobarbital on the rat liver microsomal drug-metabolizing system. Mech Ageing Dev 15:189–202
Schmucker DL, Woodhouse KW, Wang RK, Wynne H, James OF, McManus M, Kremers P (1990) Effects of age and gender on in vitro properties of human liver microsomal monooxygenases. Clin Pharmacol Ther 48:365–374
Smorenburg CH, ten Tije AJ, Verweij J, Bontenbal M, Mross K, van Zomeren DM, Seynaeve C, Sparreboom A (2003) Altered clearance of unbound paclitaxel in elderly patients with metastatic breast cancer. Eur J Cancer 39:196–202
Sotaniemi E A, Arranto AJ, Pelkonen O, Pasanen M (1997) Age and cytochrome P450-linked drug metabolism in humans: an analysis of 226 subjects with equal histopathologic conditions. Clin Pharmacol Ther 61:331–339
Stadtman ER (1992) Protein oxidation and aging. Science 257:1220–1224
Stadtman ER (2002) Importance of individuality in oxidative stress and aging. Free Radic Biol Med 33:597–604
Starke-Reed PE, Oliver CN (1989) Protein oxidation and proteolysis during aging and oxidative stress. Arch Biochem Biophys 275:559–567
Szweda PA, Friguet B, Szweda LI (2002) Proteolysis, free radicals, and aging. Free Radic Biol Med 33:29–36
Tanira MOM, Ali BH, Bashir AK (1997) Effect of endotoxin on gentamicin pharmacokinetics in old and young adult rats. Life Sci 60:413–424
Taper HS, Somer MP, Lans M, de Gerlache J, Roberfroid M (1988) Histochemical detection of the in vivo produced cellular aldehydes by means of direct Schiff’s reaction in CCl4 intoxicated rat liver. Arch Toxicol 61:406–410
Thomas PE, Bandiera S, Maines SL, Ryan DE, Levin W (1987) Regulation of cytochrome P-450j, a high affinity N-nitrosodimethylamaine demethylase, in rat hepatic microsomes. Biochemistry 26:2280–2289
van Bezooijen RL, Wang RK, Lechner MC, Schmucker DL (1994) Aging effects on hepatic NADPH cytochrome P450 reductase, CYP2B1&2, and polymeric immunoglobulin receptor mRNAs in male Fischer 344 rats. Exp Gerontol 29:187–195
Warrington JS, Poku JW, von Moltke LL, Shader RI, Harmatz JS, Greenblatt DJ (2000) Effects of age on in vitro midazolam biotransformation in male CD-1 mouse liver microsomes. J Pharmacol Exp Ther 292:1024–1031
Acknowledgements
We thank Nicole Maton and Véronique Allaeys for their excellent technical assistance. Animal care and experiments were performed according to Biosafety and Ethical rules in application in Belgium as adopted by the Bioethical Committee of the Université Catholique de Louvain.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wauthier, V., Verbeeck, R.K. & Buc Calderon, P. Age-related changes in the protein and mRNA levels of CYP2E1 and CYP3A isoforms as well as in their hepatic activities in Wistar rats. What role for oxidative stress?. Arch Toxicol 78, 131–138 (2004). https://doi.org/10.1007/s00204-003-0526-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-003-0526-z