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Cocaine-induced kidney toxicity: an in vitro study using primary cultured human proximal tubular epithelial cells

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Abstract

Renal failure resulting from cocaine abuse has been well documented, although the underlying mechanisms remain to be investigated. In the present study, primary cultured human proximal tubular epithelial cells (HPTECs) of the kidney were used to investigate its ability to metabolize cocaine, as well as the cytotoxicity induced by cocaine and its metabolites benzoylecgonine (BE), ecgonine methyl ester (EME) and norcocaine (NCOC). Gas chromatography/ion trap-mass spectrometry (GC/IT-MS) analysis of HPTECs exposed to cocaine (1 mM) for 72 h confirmed its metabolism into EME and NCOC, but not BE. EME levels increased along the exposure time to cocaine, while NCOC concentration diminished after reaching a maximum at 6 h, indicating a possible secondary metabolism for this metabolite. Cocaine promoted a concentration-dependent loss of cell viability, whereas BE and EME were found to be non-toxic to HPTECs at the tested conditions. In contrast, NCOC revealed to have higher intrinsic nephrotoxicity than the parent compound. Moreover, cocaine-induced cell death was partially reversed in the presence of ketoconazole (KTZ), a potent CYP3A inhibitor, supporting the hypothesis that NCOC may play a role in cocaine-induced nephrotoxicity. Cocaine-induced cytotoxicity was found to involve intracellular glutathione depletion at low concentrations and to induce mitochondrial damage at higher concentrations. Under the present experimental conditions, HPTECs death pathway followed an apoptotic pattern, which was evident for concentrations as low as 0.1 mM.

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Abbreviations

HPTECs:

Human proximal tubular epithelial cells

BE:

Benzoylecgonine

EME:

Ecgonine methyl ester

NCOC:

Norcocaine

GC/IT-MS:

Gas chromatography/ion trap-mass spectrometry

KTZ:

Ketoconazole

CYP450:

Cytochrome P450

AEME:

Anhydroecgonine methyl ester

CE:

Cocaethylene

PS:

Phophatidylserine

ROS:

Reactive oxygen species

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Acknowledgments

This work has been supported by Fundação para a Ciência e a Tecnologia through grant no. PEst-C/EQB/LA0006/2011.

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Authors and Affiliations

  1. REQUIMTE—Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, rua Aníbal Cunha, 164, 4099-030, Porto, Portugal

    Maria João Valente, Vânia Vilas-Boas, Renata Silva, Maria de Lourdes Bastos, Félix Carvalho, Paula Guedes de Pinho & Márcia Carvalho

  2. Department of Pathology, Portuguese Oncology Institute-Porto, Porto, Portugal

    Rui Henrique

  3. Departamento de Patologia e Imunologia Molecular, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, largo Prof. Abel Salazar, 2, 4099-003, Porto, Portugal

    Rui Henrique

  4. CEBIMED, Faculty of Health Sciences, University Fernando Pessoa, Porto, Portugal

    Márcia Carvalho

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  1. Maria João Valente
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  2. Rui Henrique
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  3. Vânia Vilas-Boas
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Correspondence to Maria João Valente or Márcia Carvalho.

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Valente, M.J., Henrique, R., Vilas-Boas, V. et al. Cocaine-induced kidney toxicity: an in vitro study using primary cultured human proximal tubular epithelial cells. Arch Toxicol 86, 249–261 (2012). https://doi.org/10.1007/s00204-011-0749-3

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