Hepatotoxicity of 3,4-methylenedioxyamphetamine and α-methyldopamine in isolated rat hepatocytes: Formation of glutathione conjugates

Márcia Carvalho, Nuno Milhazes, Fernando Remião, Fernanda Borges, Eduarda Fernandes, Francisco Amado, Terrence J. Monks, Félix Carvalho, Maria Lourdes Bastos

Research output: Contribution to journalArticlepeer-review

73 Scopus citations

Abstract

The amphetamine designer drugs 3,4-methylenedioxymethamphetamine (MDMA or "ecstasy ") and its N-demethylated analogue 3,4- methylenedioxyamphetamine (MDA or "love ") have been extensively used as recreational drugs of abuse. MDA itself is a main MDMA metabolite. MDMA abuse in humans has been associated with numerous reports of hepatocellular damage. Although MDMA undergoes extensive hepatic metabolism, the role of metabolites in MDMA-induced hepatotoxicity remains unclear. Thus, the aim of the present study was to evaluate the effects of MDA and α-methyldopamine (α-MeDA), a major metabolite of MDA, in freshly isolated rat hepatocyte suspensions. The cells were incubated with MDA or α-MeDA at final concentrations of 0.1, 0.2, 0.4, 0.8, or 1.6 mM for 3 h. The toxic effects induced following incubation of hepatocyte suspensions with these metabolites were evaluated by measuring cell viability, the extent of lipid peroxidation, levels of glutathione (GSH) and glutathione disulfide (GSSG), the formation of GSH conjugates, and the activities of GSSG reductase (GR), GSH peroxidase (GPX), and GSH S-transferase (GST). MDA induced a concentration- and time-dependent GSH depletion, but had a negligible effect on lipid peroxidation, cell viability, or on the activities of GR, GPX, and GST. In contrast, α-MeDA (1.6 mM, 3 h) induced a marked depletion of GSH accompanied by a loss on cell viability, and decreases in GR, GPX and GST activities, although no significant effect on lipid peroxidation was found. For both metabolites, GSH depletion was not accompanied by increases in GSSG levels; rather, 2-(glutathion-5-yl)- α-MeDA and 5-(glutathion-S-yl)-α-MeDA were identified by HPLC-DAD/ EC within cells incubated with MDA or α-MeDA. The results provide evidence that one of the early consequences of MDMA metabolism is a disruption of thiol homeostasis, which may result in loss of protein function and the initiation of a cascade of events leading to cellular damage.

Original languageEnglish (US)
Pages (from-to)16-24
Number of pages9
JournalArchives of Toxicology
Volume78
Issue number1
DOIs
StatePublished - Jan 2004

Keywords

  • 3,4-Methylenedioxyamphetamine
  • Glutathione conjugates
  • Rat hepatocytes
  • α-Methyldopamine

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

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