D-Amphetamine Toxicity in Freshly Isolated Rat Hepatocytes: A Possible Role of CYP3A

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D-Amphetamine Toxicity in Freshly Isolated Rat Hepatocytes: A Possible Role of CYP3A

The aim of this study was to trace D-amphetamine toxicity in isolated rat hepatocytes and to elucidate a possible involvement of CYP3A in the mechanisms of its toxicity. To this end, male Wistar rats were treated with nifedipine (5 mg kg-1i.p., 5 days), a substrate and inducer of CYP3A. Hepatocytes isolated from nifedipine-treated and control rats were incubated with D-amphetamine at a concentration of 100 μmol L-1, which was determined to be an average toxic concentration (TC50) for the compound. To evaluate the possible toxic effects of D-amphetamine on freshly isolated rat hepatocytes, we assessed the following parameters: cell viability, lactate dehydrogenase (LDH) activity, and glutathione (GSH).

The results showed that nifedipine potentiated amphetamine cytotoxicity in vitro, as follows: cell viability dropped by 65% (p>0.001), GSH by 80% (p>0.001), and LDH activity increased by 190% (p>0.001). To clarify the role of nifedipine in amphetamine cytotoxicity, we used amiodarone, a substrate and an inhibitor of CYP3A. Pre-incubation of nifedipine-treated hepatocytes with amiodarone (14 μmol L-1) significantly lowered amphetamine cytotoxicity.

Our results confirmed the toxicity of D-amphetamine in isolated rat hepatocytes and the involvement of CYP3A in its metabolism and hepatotoxicity.

Caldwell J. The metabolism of amphetamines and related stimulants in animals and man. In: Caldwell J, editor. Amphetamines and related stimulants: Chemical biological and sociological aspects. Boca Raton: CRC Press Inc.; 1980. p. 29-46.

Bach MV, Coutts RT, Baker GB. Involvement of CYP2D6 in the in vitro metabolism of amphetamine, two N-alkylamphetamines and their 4-methoxylated derivatives. Xenobiotica 1999;29:719-32.

Law MY, Slawson MH, Moody DE. Selective involvement of cytochrome P450 2D subfamily in in vivo 4-hydroxylation of amphetamine in rat. Drug Metab Dispos 2000;28:348-53.

Carvalho F, Remião F, Amado F, Domingues P, Correia AJ, Bastos ML. D-Amphetamine interaction with glutathione in freshly isolated rat hepatocytes. Chem Res Toxicol 1996;9:1031-6.

Yanada H, Oguri K, Yoshimura H. Effects of several factors on urinary excretion of methamphetamine and its metabolites in rats. Xenobiotica 1986;16:137-41.

Timbrell J. 2000. Factors affecting metabolism and disposition — Enzyme inhibition. In: Principles of Biochemical Toxicology. 3rd ed. New York (NY): Taylor and Francis; 2000. p. 164-5.

Graham RJ, Benet LZ. Antiprogestin-mediated inactivation of cytochrome P 450 CYP 3A4. Int J Exp Clin Pharmacol 1998;56:150-7.

Council of Europe. European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes. CETS No. 123, 1991 [displayed 30 May 2007] Available at http://conventions.coe.int/treaty/Commun/QueVoulezVous.asp?NT=123ECL=ENG

Vitcheva V, Mitcheva M. Effect of nifedipine on behavioral and biochemical parameters in rats after multiple morphine administration. Methods Find Exp Clin Pharmacol 2004;26:631-4.

Fau D, Berson A, Eugene D, Formently B, Fisch C, Pessayre D. Mechanism for the hepatotoxicity of the antiandrogen Nilutamide. Evidence suggesting that redox cycling of this nitroaromatic drug lead to oxidative stress in isolated hepatocytes. J Pharm Exp Ther 1992;263:69-77.

Mitcheva M, Kondeva M, Vitcheva V, Nedialkov P, Kitanov G. Effect of benzophenones from Hypericum annulatum on carbon tetrachloride-induced toxicity in freshly isolated rat hepatocytes. Redox Rep 2006;11:3-8.

Ohyama K, Nakajiama M, Suzuki M, Shimada N, Yamazaki H, Yoki T. Inhibitory effects of amiodarone and its N-deethylated metabolite on human cytochrome P 450 activities: prediction of in vivo drug interactions. Br J Clin Pharmacol 2000;49:244-53.

Bergmeyer HU, editor. Methods of enzymatic analysis. Vol 1. New York (NY) / London: Verlag Chemie — Academic Press; 1974.

Advokat C. Update on amphetamine neurotoxicity and its relevance to the treatment of ADHD. J Atten Disord 2007;11:8-16.

Carvalho F, Remião F, Soares ME, Catarino R, Queiroz G, Bastos ML. D-Amphetamine-induced hepatotoxicity: possible contribution of catecholamines and hyperthermia to the effects studied in isolated hepatocytes. Arch Toxicol 1997;71:429-36.

Jones AJ, Simpson KJ. Review article: mechanisms and management of hepatotoxicity in ecstasy (MDMA) and amphetamine intoxications. Aliment Pharmacol Ther 1999;13:129-33.

Maurer HH, Kraemer T, Springer D, Staack RF. Chemistry, pharmacology, toxicology, and hepatic metabolism of designer drugs of the amphetamine (ecstasy), piperazine, and pyrrolidinophenone types: a synopsis. Ther Drug Monit 2004;26:127-31.

Antoniou T, Tseng AL. Interactions between recreational drugs and antiretroviral agents. Ann Pharmacother 2002;36:1598-613.

Shulman A, Jagoda J, Laycock G, Kelly H. Calcium channel blocking drugs in the management of drug dependence, withdrawal and craving. A clinical pilot study with nifedipine and verapamil. Aust Fam Physician 1998;27(Suppl 1):S19-24.

Kastelova A, Koleva M, Staneva-Stoytcheva D. Changes in rat liver monooxygenase activities after administration of atenolol, nifedipine and diltiazem alone and in combination. Methods Find Exp Clin Pharmacol 2000;22:627-31.

Drocourt L, Pascussi JM, Assenat E, Fabre JM, Maurel P, Vilarem MJ. Calcium channel modulators of the dihydropyridine family are human pregnane X receptor activators and inducers of CYP 3A, CYP 2B and CYP 2C in human hepatocytes. Drug Metab Dispos 2001;29:1325-31.

Diehl KH, Hull R, Morton D, Pfizer R, Rabemampianina Y, Smith D, Vidal JM, van de Vorstenbosch C. A good practice guide to the administration of substances and removal of blood, including routs and volumes. J Appl Toxicol 2001;21:15-23.

Coleman MD. Human Drug Metabolism. West Sussex: John Wiley and Sons; 2006.

Ha HR, Bigler L, Binder M, Korlik P, Stieger B, Heese M, Altorfer HR, Follath F. Metabolism of amiodarone (part I): identification of a new hydroxylated metabolite of amiodarone. Drug Metab Dispos 2001;29:152-8.

Archives of Industrial Hygiene and Toxicology

The Journal of Institute for Medical Research and Occupational Health

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