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Guanosine prevents nitroxidative stress and recovers mitochondrial membrane potential disruption in hippocampal slices subjected to oxygen/glucose deprivation

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Abstract

Guanosine, the endogenous guanine nucleoside, prevents cellular death induced by ischemic events and is a promising neuroprotective agent. During an ischemic event, nitric oxide has been reported to either cause or prevent cell death. Our aim was to evaluate the neuroprotective effects of guanosine against oxidative damage in hippocampal slices subjected to an in vitro ischemia model, the oxygen/glucose deprivation (OGD) protocol. We also assessed the participation of nitric oxide synthase (NOS) enzymes activity on the neuroprotection promoted by guanosine. Here, we showed that guanosine prevented the increase in ROS, nitric oxide, and peroxynitrite production induced by OGD. Moreover, guanosine prevented the loss of mitochondrial membrane potential in hippocampal slices subjected to OGD. Guanosine did not present an antioxidant effect per se. The protective effects of guanosine were mimicked by inhibition of neuronal NOS, but not of inducible NOS. The neuroprotective effect of guanosine may involve activation of cellular mechanisms that prevent the increase in nitric oxide production, possibly via neuronal NOS.

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Abbreviations

1400 W:

N-(3-(Aminomethyl)benzyl)acetamidine

7-NI:

7-nitroindazole

eNOS:

Endothelial nitric oxide synthase

GUO:

Guanosine

HBSS:

Hank’s balanced salt solution

iNOS:

Inducible nitric oxide synthase

KRB:

Krebs-Ringer bicarbonate buffer

L-NAME:

L-N G-nitro-L-arginine methyl ester

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

nNOS:

Neuronal nitric oxide synthase

OGD:

Oxygen/glucose deprivation

ONOO− :

Peroxynitrite

ROS:

reactive oxygen species

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Acknowledgments

Research supported by grants from the Brazilian funding agencies to C.I.T.: CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico)–Projects IBN-Net # 01.06.0842-00 and INCT for Excitotoxicity and Neuroprotection; CAPES (Coordenação do Pessoal de Ensino Superior)–Project CAPES-PVE 052/2012; FAPESC (Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina)–Project NENASC. C.I.T. is a recipient of CNPq productivity fellowship and M.P.C. is a recipient of CAPES/PNPD post-doctoral scholarship. The authors state no conflicts of interest. All authors have materially participated in the research and/or article preparation.

Compliance with ethical standards

The procedures used in the present study complied with the guidelines on animal care of the UFSC Ethics Committee on the Use of Animals (CEUA), which follows the “Principles of laboratory animal care” from NIH (2011).

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Authors and Affiliations

  1. Departamento de Bioquímica, CCB, UFSC, Universidade Federal de Santa Catarina, Trindade, 88040-900, Florianópolis, SC, Brazil

    Andreza F. de Bem & Carla I. Tasca

  2. Programa de Pós-Graduação em Bioquímica, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Daniel T. Thomaz, Maurício Peña Cunha, Andreza F. de Bem & Carla I. Tasca

  3. Programa de Pós-Graduação em Neurociências, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Florianópolis, SC, Brazil

    Tharine A. Dal-Cim, Wagner C. Martins, Débora Lanznaster, Andreza F. de Bem & Carla I. Tasca

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  1. Daniel T. Thomaz
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  2. Tharine A. Dal-Cim
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Correspondence to Carla I. Tasca.

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Thomaz, D.T., Dal-Cim, T.A., Martins, W.C. et al. Guanosine prevents nitroxidative stress and recovers mitochondrial membrane potential disruption in hippocampal slices subjected to oxygen/glucose deprivation. Purinergic Signalling 12, 707–718 (2016). https://doi.org/10.1007/s11302-016-9534-3

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