Evaluating the Role of Drug Metabolism and Reactive Intermediates in Trazodone-Induced Cytotoxicity toward Freshly-Isolated Rat Hepatocytes

Abstract

Background: Trazodone is an antidepressant agent widely administered for the treatment of depressive disorders. On the other hand, several cases of hepatic injury have been reported after Trazodone administration. Although the precise mechanism(s) of trazodone-induced liver injury is not known, some investigations proposed the role of reactive intermediates in this complication. This study was designed to investigate the role of reactive metabolites in hepatocytes injury induced by trazodone.

Methods: Isolated rat hepatocytes were prepared by the method of collagenase enzyme perfusion via the portal vein. Cells were treated with trazodone, its cytotoxic metabolite, and different enzyme inhibitors and cytoprotective agents.

Results: It was found that trazodone was toxic towards hepatocytes and caused 50% cell death after 2 h of incubation at a dose of 450 µM. The trazodone postulated reactive metabolite; m-chlorophenyl piperazine (m-CPP) was less toxic and caused 50% cell death at a dose of 750 µM at a similar time period. Cellular glutathione (GSH) depletion and lipid peroxidation were detected when hepatocytes were treated with trazodone and/or m-CPP. Depleting hepatocytes GSH beforehand, increased cytotoxicity of both trazodone and m-CPP. Troleandomycin as the CYP3A4 inhibitor prevented cytotoxicity of trazodone but slightly affected m-CPP-induced cell injury. Inhibition of CYP2D6 by quinidine and cimetidine increased the cytotoxicity of both trazodone and m-CPP. Antioxidants and ATP suppliers slightly prevented cytotoxicity of trazodone and m-CPP.

Conclusion: As inhibitors of CYP3A4 and 2D6 affected trazodone cytotoxicity, it is suggested that trazodone -induced cytotoxicity, at least in part, is mediated by its reactive metabolites.

Key words

drug hepatotoxicity - glutathione - hepatocyte - oxidative stress - reactive metabolites

• References

• 1 Brogden RN, Heel RC, Speight TM et al. Trazodone: a review of its pharmacological properties and therapeutic use in depression and anxiety. Drugs 1981; 21: 401-429
• 2 Kalgutkar AS, Henne KR, Lame ME et al. Hop CECA. Metabolic activation of the nontricyclic antidepressant trazodone to electrophilic quinone-imine and epoxide intermediates in human liver microsomes and recombinant P4503A4. Chem Biol Interact 2005; 155: 10-20
• 3 Odagaki Y, Toyoshima R, Yamauchi T. Trazodone and its active metabolite m-chlorophenylpiperazine as partial agonists at 5-HT1A receptors assessed by [35S] GTPγS binding. J Psychopharm 2005; 19: 235-241
• 4 Rotzinger S, Fang J, Coutts RT et al. Human CYP2D6 and metabolism of m-chlorophenylpiperazine. Biol Psychiatry 1998; 44: 1185-1191
• 5 Wen B, Ma L, Rodrigues AD et al. Detection of novel reactive metabolites of trazodone: evidence for CYP2D6-mediated bioactivation of m-chlorophenylpiperazine. Drug Metab Disposition 2008; 36: 841-850
• 6 Maes M, Westenberg H, Vandoolaeghe E et al. Effects of trazodone and fluoxetine in the treatment of major depression: therapeutic pharmacokinetic and pharmacodynamic interactions through formation of meta-chlorophenylpiperazine. J Clin Psychopharmacol 1997; 17: 358-364
• 7 Yasui N, Otani K, Kaneko S et al. Inhibition of trazodone metabolism by thioridazine in humans. Ther Drug Monit 1995; 17: 333-335
• 8 Otani K, Ishida M, Kaneko S et al. Effects of carbamazepine coadministration on plasma concentrations of trazodone and its active metabolite, m-chlorophenylpiperazine. Ther Drug Monit 1996; 18: 164-167
• 9 Rettman KS, McClintock C. Hepatotoxicity after short-term trazodone therapy. The Annals of pharmacotherapy 2001; 35: 1559-1561
• 10 Fernandes NF, Martin RR, Schenker S. Trazodone-induced hepatotoxicity: a case report with comments on drug-induced hepatotoxicity. The American journal of gastroenterology 2000; 95: 532-535
• 11 DeSanty KP, Amabile CM. Antidepressant-induced liver injury. Ann Pharmacother 2007; 41: 1201-1211
• 12 Taziki S, Sattari MR, Eghbal MA. Mechanisms of trazodone-induced cytotoxicity and the protective effects of melatonin and/or taurine toward freshly isolated rat hepatocytes. J Biochem Mol Toxicol 2013; 27: 457-462
• 13 Eghbal MA, Taziki S, Sattari MR. Mechanisms of phenytoin-induced toxicity in freshly isolated rat hepatocytes and the protective effects of taurine and/or melatonin. J Biochem Mol Toxicol 2014; 28: 111-118
• 14 O'Brien PJ, Siraki AG. Accelerated cytotoxicity mechanism screening using drug metabolising enzyme modulators. Curr Drug Metab 2005; 6: 101-109
• 15 Nafisi S, Heidari R, Ghaffarzadeh M et al. Cytoprotective effects of silafibrate, a newly-synthesised siliconated derivative of clofibrate, against acetaminophen-induced toxicity in isolated rat hepatocytes. Arh Hig Rada Toksikol 2014; 65: 169-178
• 16 Heidari R, Babaei H, Eghbal MA. Amodiaquine-induced toxicity in isolated rat hepatocytes and the cytoprotective effects of taurine and/or N-acetyl cysteine. Res Pharm Sci 2014; 9: 97-105
• 17 Heidari R, Babaei H, Eghbal MA. Cytoprotective Effects of Organosulfur Compounds against Methimazole Induced Toxicity in Isolated Rat Hepatocytes. Adv Pharm Bull 2013; 3: 135-142
• 18 Heidari R, Babaei H, Eghbal MA. Ameliorative effects of taurine against methimazole-induced cytotoxicity in isolated rat hepatocytes. Sci Pharm 2012; 80: 987-999
• 19 Heidari R, Babaei H, Eghbal M. Mechanisms of methimazole cytotoxicity in isolated rat hepatocytes. Drug Chem Toxicol 2013; 36: 403-411
• 20 Vandeputte C, Guizon I, Genestie-Denis I et al. A microtiter plate assay for total glutathione and glutathione disulfide contents in cultured/isolated cells: performance study of a new miniaturized protocol. Cell Biol Toxicol 1994; 10: 415-421
• 21 Abdoli N, Heidari R, Azarmi Y et al. Mechanisms of the statins cytotoxicity in freshly isolated rat hepatocytes. J Biochem Mol Toxicol 2013; 27: 287-294
• 22 Khan S, O’Brien PJ. 1-bromoalkanes as new potent nontoxic glutathione depletors in isolated rat hepatocytes. Biochem Biophys Res Commun 1991; 179: 436-441
• 23 O’Brien PJ, Siraki AG. Accelerated cytotoxicity mechanism screening using drug metabolising enzyme modulators. Curr Drug Metab 2005; 6: 101-109
• 24 Jamshidzadeh A, Niknahad H, Kashafi H. Cytotoxicity of chloroquine in isolated rat hepatocytes. Journal of Applied Toxicology 2007; 27: 322-326
• 25 Heidari R, Babaei H, Eghbal MA. Cytoprotective effects of taurine against toxicity induced by isoniazid and hydrazine in isolated rat hepatocytes. Archives of Industrial Hygiene and Toxicology 2013; 64: 201-210
• 26 Kim D-O, Lee CY. Comprehensive study on vitamin C equivalent antioxidant capacity (VCEAC) of various polyphenolics in scavenging a free radical and its structural relationship. Crit Rev Food Sci Nutr 2004; 44: 253-273
• 27 Rotzinger S, Fang J, Baker GB. Trazodone is metabolized tom-Chlorophenylpiperazine by CYP3A4 from human sources. Drug Metab Disposition 1998; 26: 572-575
• 28 Andrade RJ, Agundez JA, Lucena MI et al. Pharmacogenomics in drug induced liver injury. Curr Drug Metab 2009; 10: 956-970
• 29 Kim SH, Kim SH, Bahn JW et al. Genetic polymorphisms of drug-metabolizing enzymes and anti-TB drug-induced hepatitis. Pharmacogenomics 2009; 10: 1767-1779
• 30 Heidari R, Taheri V, Rahimi HR et al. Sulfasalazine-induced renal injury in rats and the protective role of thiol-reductants. Renal Failure 2015; 38: 137-141