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Neuronal Nitric Oxide Synthase is a Key Factor in Doxorubicin-Induced Toxicity to Rat-Isolated Cortical Neurons

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Abstract

Doxorubicin (DOX) is neurotoxic to serum-free cultures of rat cortical neurons in a biphasic concentration manner. For concentrations up to 0.5 μM, cell death follows an apoptotic pattern, while for higher concentrations apoptosis is inhibited and necrosis becomes dominant. Considering the potential toxic effects of DOX resulting from its redox-cycling, in this study we investigated the generation of reactive species and subsequent oxidative stress effects, formation of quinoproteins, activation of NF-kB, depletion of energy levels and consequent cell death in cultures of primary rat cortical cells challenged with this antitumour drug. The influence of neuronal nitric oxide synthase (nNOS) on DOX-induced neuronal cell damage was subsequently evaluated. The exposure of rat cortical primary cell cultures to DOX resulted in a significant generation of ROS/RNS, activation of NF-kB, depletion of GSH levels, depletion of ATP, and cell death, in a concentration biphasic manner. Doxorubicin also significantly increased protein-bound quinone products in neurons in a concentration-dependent manner. Inhibition of nNOS decreased neuronal cell death induced by DOX in a significant way, at the first phase of the biphasic curve. In conclusion, this study shows, for the first time, a clear involvement of nNOS and subsequent ROS/RNS generation as crucial signalling mediators of DOX-induced neurotoxicity on isolated cortical neurons. Inhibition of ROS/RNS formation, modulation of NOS isoforms and modulation of NF-kB activation could be of beneficial in preventing damage in the CNS caused by DOX.

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Acknowledgements

The authors would like to thank Professor Ulrich Dirnagl for his help in the integration and discussion of the presented data. Miguel Ângelo Lopes was the recipient of a PhD grant from “Fundação para a Ciência e Tecnologia” (Ref. SFRD/BD/17899/2004).

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Authors and Affiliations

  1. REQUIMTE, Faculty of Pharmacy, Toxicology Department, University of Porto, Rua Aníbal Cunha 164, 4050-047, Porto, Portugal

    Miguel Ângelo Lopes, Félix Dias Carvalho & Maria de Lourdes Bastos

  2. Department of Experimental Neurology, Center for Stroke Research, Charité-Universitätsmedizin, Berlin, Germany

    Miguel Ângelo Lopes & Andreas Meisel

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  1. Miguel Ângelo Lopes
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  2. Andreas Meisel
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  3. Félix Dias Carvalho
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  4. Maria de Lourdes Bastos
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Correspondence to Miguel Ângelo Lopes or Maria de Lourdes Bastos.

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Lopes, M.Â., Meisel, A., Carvalho, F.D. et al. Neuronal Nitric Oxide Synthase is a Key Factor in Doxorubicin-Induced Toxicity to Rat-Isolated Cortical Neurons. Neurotox Res 19, 14–22 (2011). https://doi.org/10.1007/s12640-009-9135-9

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  • DOI: https://doi.org/10.1007/s12640-009-9135-9

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