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Guanosine Exerts Neuroprotective Effect in an Experimental Model of Acute Ammonia Intoxication

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Abstract

The nucleoside guanosine (GUO) increases glutamate uptake by astrocytes and acts as antioxidant, thereby providing neuroprotection against glutamatergic excitotoxicity, as we have recently demonstrated in an animal model of chronic hepatic encephalopathy. Here, we investigated the neuroprotective effect of GUO in an acute ammonia intoxication model. Adult male Wistar rats received an intraperitoneal (i.p.) injection of vehicle or GUO 60 mg/kg, followed 20 min later by an i.p. injection of vehicle or 550 mg/kg of ammonium acetate. Afterwards, animals were observed for 45 min, being evaluated as normal, coma (i.e., absence of corneal reflex), or death status. In a second cohort of rats, video-electroencephalogram (EEG) recordings were performed. In a third cohort of rats, the following were measured: (i) plasma levels of glucose, transaminases, and urea; (ii) cerebrospinal fluid (CSF) levels of ammonia, glutamine, glutamate, and alanine; (iii) glutamate uptake in brain slices; and (iv) brain redox status and glutamine synthetase activity in cerebral cortex. GUO drastically reduced the lethality rate and the duration of coma. Animals treated with GUO had improved EEG traces, decreased CSF levels of glutamate and alanine, lowered oxidative stress in the cerebral cortex, and increased glutamate uptake by astrocytes in brain slices compared with animals that received vehicle prior to ammonium acetate administration. This study provides new evidence on mechanisms of guanine-derived purines in their potential modulation of glutamatergic system, contributing to GUO neuroprotective effects in a rodent model of by acute ammonia intoxication.

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Acknowledgments

This work was supported by Brazilian agencies and grants: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS), and Brazilian Institute of Neuroscience—IBN Net FINEP, INCT—Excitotoxicity and Neuroprotection (573577/2008-5).

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    Authors and Affiliations

    1. Postgraduate Program in Biological Sciences: Biochemistry, ICBS, Federal University of Rio Grande do Sul, Porto Alegre, RS, 90035-003, Brazil

      G. F. Cittolin-Santos, A. M. de Assis, P. A. Guazzelli, L. G. Paniz, J. S. da Silva, M. E. Calcagnotto, G. Hansel, K. C. Zenki, E. Kalinine & D. O. Souza

    2. Department of Biochemistry, Federal University of Rio Grande do Sul, Porto Alegre, RS, 90035-003, Brazil

      M. E. Calcagnotto & D. O. Souza

    3. Department of Physiology, Federal University of Sergipe, São Cristovão, SE, 49100-000, Brazil

      K. C. Zenki & E. Kalinine

    4. Health Sciences Center, Lutheran University of Brazil (ULBRA), Campus Santa Maria, Santa Maria, RS, 97020-001, Brazil

      M. M. Duarte

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    1. G. F. Cittolin-Santos
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    2. A. M. de Assis
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    Correspondence to D. O. Souza.

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    All experiments were approved by the Ethics Commission (CEUA/UFRGS) under project number 29468 and followed the National Institutes of Health “Guide for the Care and Use of Laboratory Animals” (NIH publication No. 80-23, revised 1996).

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    The authors declare that they have no competing financial interests.

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    G. F. Cittolin-Santos and A. M. de Assis contributed equally to this work.

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    Cittolin-Santos, G.F., de Assis, A.M., Guazzelli, P.A. et al. Guanosine Exerts Neuroprotective Effect in an Experimental Model of Acute Ammonia Intoxication. Mol Neurobiol 54, 3137–3148 (2017). https://doi.org/10.1007/s12035-016-9892-4

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