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Interferon Gamma Potentiates the Injury Caused by MPP(+) on SH-SY5Y Cells, Which is Attenuated by the Nitric Oxide Synthases Inhibition

Neurochemical Research volume 39, pages 2452–2464 (2014)Cite this article

Abstract

This study examined whether the cytokine interferon (IFN) gamma plays a role in the injury of SH-SY5Y cells caused by MPP(+) (1-methyl-4-phenylpyridinium). First of all, IFN-gamma sensitized cells to the neurotoxin MPP(+), as determined by MTT (3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide) assay. MPP(+)-injured cells showed higher reactive oxygen species (ROS) levels, which was reinforced by IFN-gamma. The injury triggered a marked expression of the neuronal NOS (nNOS) enzyme. L-NAME [N(ω)-nitro-l-arginine methyl ester, a non-specific NOS inhibitor] reestablished the cell viability after IFN-gamma challenging, and recovered cells from MPP(+) injury (95.0 vs. 84.7 %; P < 0.05). Seven-NI (7-nitroindazole, a nNOS inhibitor) protected cells against the injury by MPP(+) co-administered with IFN-gamma. Both inhibitors restrained the apoptosis of SH-SY5Y cells caused by MPP(+)/IFN-gamma. Regarding oxidative stress, L-NAME and 7-NI attenuated the increase in ROS levels caused by MPP(+) (45.3 or 48.4 vs. 87.9 %, P < 0.05). Indeed, L-NAME was more effective than 7-NI for reducing oxidative stress caused by MPP(+) under IFN-gamma exposition. The nNOS gene silencing by small-interfering RNAs recovered cells challenged by IFN-gamma (24 h), or MPP(+) (8 h). In conclusion, IFN-gamma sensitizes cells to MPP(+)-induced injury, also causing an increase in ROS levels. Pretreating cells with L-NAME or 7-NI reverts both the oxidative stress and apoptosis triggered by the neurotoxin MPP(+). Taking together, our data reinforce that IFN-gamma and NOS enzymes play a role in oxidative stress and dopaminergic cell death triggered by MPP(+).

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Acknowledgments

The present research received financial support from the following Brazilian institutions: Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES, Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, and Fundação de Apoio à Pesquisa do Distrito Federal—FAP-DF.

Conflict of interest

Data regarding nNOS sequences used for designing the structures siRNAnNOShum_3987 (SEQ ID NO: 38/39) and siRNAnNOShum_4400 (SEQ ID NO: 41/42) are under patent registration at the Brazilian Institute for Industrial Property—“Instituto Nacional da Propriedade Industrial”—INPI, BR 10 2012 032844 5.

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  1. Laboratório de Tecnologias para Terapia Gênica, ASS 128, ICC Sul. Universidade de Brasília - UnB, Campus Darcy Ribeiro, FAV, Brasília, DF, 70910-970, Brazil

    Simoneide S. Titze-de-Almeida, Cátia Faria Lustosa, Camila Hillesheim Horst & Ricardo Titze-de-Almeida

  2. Laboratório de Neurofisiologia e Biologia Molecular, Dept. Morfologia Fisiologia e Patologia Básica, FORP, Universidade de São Paulo - USP, Ribeirão Prêto, SP, 14040-904, Brazil

    Elaine Del Bel

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  1. Simoneide S. Titze-de-Almeida
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Titze-de-Almeida, S.S., Lustosa, C.F., Horst, C.H. et al. Interferon Gamma Potentiates the Injury Caused by MPP(+) on SH-SY5Y Cells, Which is Attenuated by the Nitric Oxide Synthases Inhibition. Neurochem Res 39, 2452–2464 (2014). https://doi.org/10.1007/s11064-014-1449-1

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Keywords

  • SH-SY5Y
  • Neurodegeneration
  • Nitric oxide synthases
  • Oxidative stress
  • Interferon gamma
  • MPP(+)