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"Ecstasy"-induced toxicity in SH-SY5Y differentiated cells: role of hyperthermia and metabolites

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Abstract

3,4-Methylenedioxymethamphetamine (MDMA; “ecstasy”) is a recreational hallucinogenic drug of abuse known to elicit neurotoxic properties. Hepatic formation of neurotoxic metabolites is thought to play a major role in MDMA-related neurotoxicity, though the mechanisms involved are still unclear. Here, we studied the neurotoxicity mechanisms and stability of MDMA and 6 of its major human metabolites, namely α-methyldopamine (α-MeDA) and N-methyl-α-methyldopamine (N-Me-α-MeDA) and their correspondent glutathione (GSH) and N-acetyl-cysteine (NAC) conjugates, under normothermic (37 °C) or hyperthermic conditions (40 °C), using cultured SH-SY5Y differentiated cells. We showed that MDMA metabolites exhibited toxicity to SH-SY5Y differentiated cells, being the GSH and NAC conjugates more toxic than their catecholic precursors and MDMA. Furthermore, whereas the toxicity of the catechol metabolites was potentiated by hyperthermia, NAC-conjugated metabolites revealed higher toxicity under normothermia and GSH-conjugated metabolites-induced toxicity was temperature-independent. Moreover, a time-dependent decrease in extracellular concentration of MDMA metabolites was observed, which was potentiated by hyperthermia. The antioxidant NAC significantly protected against the neurotoxic effects of MDMA metabolites. MDMA metabolites increased intracellular glutathione levels, though depletion in thiol content was observed in MDMA-exposed cells. Finally, the neurotoxic effects induced by the MDMA metabolite N-Me-α-MeDA involved caspase 3 activation. In conclusion, this study evaluated the stability of MDMA metabolites in vitro, and demonstrated that the catechol MDMA metabolites and their GSH and NAC conjugates, rather than MDMA itself, exhibited neurotoxic actions in SH-SY5Y differentiated cells, which were differently affected by hyperthermia, thus highlighting a major role for reactive metabolites and hyperthermia in MDMA’s neurotoxicity.

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Acknowledgments

This work was supported by “Fundação para a Ciência e a Tecnologia (FCT),” Portugal (Project PTDC/SAU-FCF/102958/2008), under the framework of “Programa Operacional Temático Factores de Competitividade (COMPTE) do Quadro Comunitário de Apoio III” and “Fundo Comunitário Europeu (FEDER) (FCOMP-01-0124-FEDER-011079).” The NMR spectrometers are part of the National NMR Facility, supported by FCT (RECI/BBB-BQB/0230/2012). DJB was supported by a fellowship (SFRH/BD/64939/2009) from FCT, Portugal.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. REQUIMTE (Rede de Química e Tecnologia), Toxicology Laboratory, Department of Biological Sciences, Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira 228, 4050-313, Porto, Portugal

    Daniel José Barbosa, João Paulo Capela, Renata Silva, Maria Lourdes Bastos & Félix Carvalho

  2. Faculty of Health Sciences, University Fernando Pessoa, Rua Carlos da Maia 296, 4200-150, Porto, Portugal

    João Paulo Capela

  3. REQUIMTE/CQFB (Centro de Química Fina e Biotecnologia), Department of Chemistry, Faculty of Science and Technology, University Nova de Lisboa, 2829-516, Caparica, Portugal

    Luísa Maria Ferreira & Paula Sério Branco

  4. REQUIMTE, Laboratory of Applied Chemistry, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Rua Jorge Viterbo Ferreira 228, 4050-313, Porto, Portugal

    Eduarda Fernandes

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  1. Daniel José Barbosa
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  2. João Paulo Capela
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  3. Renata Silva
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  4. Luísa Maria Ferreira
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Correspondence to Daniel José Barbosa or Félix Carvalho.

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Barbosa, D.J., Capela, J.P., Silva, R. et al. "Ecstasy"-induced toxicity in SH-SY5Y differentiated cells: role of hyperthermia and metabolites. Arch Toxicol 88, 515–531 (2014). https://doi.org/10.1007/s00204-013-1147-9

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