Effect of Alteration of Glutathione Content on Cell Viability in α-Synuclein-Transfected SH-SY5Y Cells

Ken-Ichi  Tanaka; Kanako  Sonoda; Masato  Asanuma

doi:10.4236/apd.2017.63010

Ken-Ichi Tanaka¹^*, Kanako Sonoda², Masato Asanuma²

Abstract: It is well known that α-synuclein (αS) plays an important role in the pathogenesis of Parkinson’s disease (PD). Moreover, oxidative stress is also thought to be an important factor in PD due to induction of dopaminergic neuronal cell death by free radicals and enhancement of αS fibrillation by oxidized stress. In the present study, to clarify the role of glutathione (GSH), an intracellular antioxidant, on the molecular mechanism of αS-induced cell injury, we examined the effects of L-buthionine-SR-sulfoximine (BSO), a GSH synthase inhibitor, with or without N-acetyl-L-cysteine (NAC), a source of GSH, on αS-induced cell injury in human neuroblastoma SH-SY5Y cells. Treatment with BSO significantly reduced the cell viability of both empty-vector- and αS-transfected SH-SY5Y cells in a dose-dependent manner (p < 0.01), although the ratio of αS-induced reduction of cell viability in α-syn-transfected cells was much greater than that in empty-vector-transfected cells. Moreover, BSO significantly reduced the intracellular total GSH level in both types of transformant cells. However, NAC significantly prevented BSO-induced reduction of both cell viability and GSH level in the αS-transfected cells. These findings suggest that GSH plays an important role in αS-induced cell injury by reducing cell viability.

Keywords: α-Synuclein, L-Buthionine-SR-Sulfoximine, N-Acetyl-L-Cysteine, Glutathione, SH-SY5Y Cells, Parkinson’s Disease

Cite this paper: Tanaka, K. , Sonoda, K. and Asanuma, M. (2017) Effect of Alteration of Glutathione Content on Cell Viability in α-Synuclein-Transfected SH-SY5Y Cells. Advances in Parkinson's Disease, 6, 93-100. doi: 10.4236/apd.2017.63010.

References

[1] Eriksen, J.L., Wszolek, Z. and Petrucelli, L. (2005) Molecular Pathogenesis of Parkinson Disease. Archives of Neurology, 62, 353-357. https://doi.org/10.1001/archneur.62.3.353

[2] Dehay, B., Bourdenx, M., Gorry, P., Przedborski, S., Vila, M., Hunot, S., Singleton, A., Olanow, C.W., Merchant, K.M., Bezard, E., Petsko, G.A. and Meissner, W.G. (2015) Targeting α-Synuclein for Treatment of Parkinson’s Disease: Mechanistic and Therapeutic Considerations. The Lancet Neurology, 14, 855-866. https://doi.org/10.1016/S1474-4422(15)00006-X

[3] Roberts, H.L. and Brown, D.R. (2015) Seeking a Mechanism for the Toxicity of Oligomeric α-Synuclein. Biomolecules, 5, 282-305. https://doi.org/10.3390/biom5020282

[4] Sulzer, D., Bogulavsky, J., Larsen, K.E., Behr, G., Karatekin, E., Kleinman, M.H., Turro, N., Krantz, D., Edwards, R.H., Greene, L.A. and Zecca, L. (2000) Neuromelanin Biosynthesis Is Driven by Excess Cytosolic Catecholamines Not Accumulated by Synaptic Vesicles. Proceedings of the National Academy of Sciences of the USA, 97, 11869-11874.
https://doi.org/10.1073/pnas.97.22.11869

[5] Graham, D.G. (1978) Oxidative Pathway for Catecholamines in the Genesis of Neuromelanin and Cytotoxic Quinones. Molecular Pharmacology, 14, 633-643.

[6] Dias, V., Junn, E. and Mouradian, M.M. (2013) The Role of Oxidative Stress in Parkinson’s Disease. Journal of Parkinson’s Disease, 3, 461-491.

[7] Miyazaki, I. and Asanuma, M. (2008) Dopaminergic Neuron-Specific Oxidative Stress Caused by Dopamine Itself. Acta Medica Okayama, 62, 141-150.

[8] Cooper, A.J.L. and Kristal, B.S. (1997) Multiple Roles of Glutathione in the Central Nervous System. Biological Chemistry, 378, 793-802.

[9] Hall, A. (1999) The Role of Glutathione in the Regulation of Apoptosis. European Journal of Clinical Investigation, 29, 238-245. https://doi.org/10.1046/j.1365-2362.1999.00447.x

[10] Asanuma, M., Miyazaki, I., Diaz-Corrales, F.J. and Ogawa, N. (2004) Quinone Formation as Dopaminergic Neuron-Specific Oxidative Stress in the Pathogenesis of Sporadic Parkinson’s Disease and Neurotoxin-Induced Parkinsonism. Acta Medica Okayama, 58, 221-233.

[11] Higashi, Y., Asanuma, M., Miyazaki, I., Haque, M.E., Fujita, N., Tanaka, K. and Ogawa, N. (2002) The p53-Activated Gene, PAG608, Requires a Zinc Finger Domain for Nuclear Localization and Oxidative Stress-Induced Apoptosis. Journal of Biological Chemistry, 277, 42224-42232. https://doi.org/10.1074/jbc.M203594200

[12] Tanaka, K., Fujita, N., Higashi, Y. and Ogawa, N. (2002) Neuroprotective and Antioxidant Properties of FKBP-Binding Immunophilin Ligands Are Independent on the FKBP12 Pathway in Human Cells. Neuroscience Letters, 330, 147-150.

[13] Clark, J., Clore, E.L., Zheng, K., Adame, A., Masliah, E. and Simon, D.K. (2010) Oral N-Acetyl-Cysteine Attenuates Loss of Dopaminergic Terminals in α Synuclein Overexpressing Mice. PLOS One, 5, 1-10. https://doi.org/10.1371/journal.pone.0012333

[14] Dawson, T.M. and Dawson, V.L. (2003) Molecular Pathway of Neurodegeneration in Parkinson’s Disease. Science, 302, 819-822. https://doi.org/10.1126/science.1087753

[15] Scarlata, S. and Golebiewska, U. (2014) Linking Alpha-Synuclein Properties with Oxidation: A Hypothesis on a Mechanism Underling Cellular Aggregation. Journal of Bioenergetics and Biomembranes, 46, 93-98. https://doi.org/10.1007/s10863-014-9540-5

[16] Dringen, R. and Hamprecht, B. (1999) N-Acetylcysteine, But Not Methione or 2-Oxothiazolidine-4-Carboxylate, Serves as Cysteine Donor for the Synthesis of Glutathione in Cultured Neurons Derived from Embryonal Rat Brain. Neuroscience Letters, 259, 79-82.

[17] Banaclocha, M.M. (2000) N-Acetylcysteine Elicited Increase in Complex I Activity in Synaptic Mitochondria from Aged Mice: Implications for Treatment of Parkinson’s Disease. Brain Research, 859, 173-175.

[18] Xu, B., Wu, S.W., Lu, C.W., Deng, Y., Liu, W., Wei, Y.G., Yang, T.Y. and Xu, Z.F. (2013) Oxidative Stress Involvement in Manganese-Induced Alpha-Synuclein Oligomerization in Organotypic Brain Slice Cultures. Toxicology, 305, 71-78.