Suppression of MAPK attenuates neuronal cell death induced by activated glia-conditioned medium in alpha-synuclein overexpressing SH-SY5Y cells

Journal of Neuroinflammation

Cristoforo Scavone²  Maria Christina W. Avellar¹  Andrea R. Vasconcelos²  Alexandre Denadai-Souza¹  Lidia M. Yshii² 

[1] Department of Pharmacology, Section of Experimental Endocrinology, Federal University of São Paulo, São Paulo, Brazil;Department of Pharmacology, Molecular Neuropharmacology Laboratory, Institute of Biomedical Science ICB-1, University of São Paulo, Avenida Professor Lineu Prestes, 1524, São Paulo 05508-900, Brazil

关键词: Cell death;    NF-κB;    MAPK;    α-Synuclein;   

Others  :  1230498
DOI  :  10.1186/s12974-015-0412-7

 received in 2015-06-02, accepted in 2015-10-16,  发布年份 2015

【 摘 要 】

Background

Parkinson’s disease (PD) is a neurodegenerative disease with characteristics and symptoms that are well defined. Nevertheless, its aetiology remains unknown. PD is characterized by the presence of Lewy bodies inside neurons. α-Synuclein (α-syn) is a soluble protein present in the pre-synaptic terminal of neurons. Evidence suggests that α-syn has a fundamental role in PD pathogenesis, given that it is an important component of Lewy bodies localized in the dopaminergic neurons of PD patients.

Methods

In the present study, we investigated the influence of wild type (WT) and A30P α-syn overexpression on neuroblastoma SH-SY5Y toxicity induced by the conditioned medium (CM) from primary cultures of glia challenged with lipopolysaccharide (LPS) from Escherichia coli.

Results

We observed that SH-SY5Y cells transduced with α-syn (WT or A30P) and treated with CM from LPS-activated glia cells show evidence of cell death, which is not reverted by NF-κB inhibition by sodium salicylate or by blockage of P50 (NF-κB subunit). Furthermore, the expression of A30P α-syn in neuroblastoma SH-SY5Y decreases the cell death triggered by the CM of activated glia versus WT α-syn or control group. This effect of A30P α-syn may be due to the low MAPK42/44 phosphorylation. This finding is substantiated by MEK1 inhibition by PD98059, decreasing LDH release by CM in SH-SY5Y cells.

Conclusion

Our results suggest that SH-SY5Y cells transduced with α-syn (WT or A30P) and treated with CM from LPS-activated glia cells show cell death, which is not reverted by NF-κB blockage. Additionally, the expression of A30P α-syn on neuroblastoma SH-SY5Y leads to decreased cell death triggered by the CM of activated glia, when compared to WT α-syn or control group. The mechanism underlying this process remains to be completely elucidated, but the present data suggest that MAPK42/44 phosphorylation plays an important role in this process.

Trial Registration

PROSPERO: CRD42015020829

【 授权许可】

   
2015 Yshii et al.

【 图 表 】

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

【 参考文献 】

• [1]Lees AJ, Hardy J, Revesz T: Parkinson’s disease. Lancet 2009, 373:2055-2066.
• [2]Bartus RT, Baumann TL, Siffert J, Herzog CD, Alterman R, Boulis N, Turner DA, Stacy M, Lang AE, Lozano AM, Olanow CW: Safety/feasibility of targeting the substantia nigra with AAV2-neurturin in Parkinson patients. Neurology 2013, 80:1698-1701.
• [3]Dauer W, Przedborski S: Parkinson’s disease: mechanisms and models. Neuron 2003, 39:889-909.
• [4]Spillantini MG, Crowther RA, Jakes R, Hasegawa M, Goedert M: alpha-Synuclein in filamentous inclusions of Lewy bodies from Parkinson’s disease and dementia with Lewy bodies. Proc Natl Acad Sci U S A 1998, 95:6469-6473.
• [5]Spillantini MG, Schmidt ML, Lee VM, Trojanowski JQ, Jakes R, Goedert M: Alpha-synuclein in Lewy bodies. Nature 1997, 388:839-840.
• [6]Fortin DL, Troyer MD, Nakamura K, Kubo S, Anthony MD, Edwards RH: Lipid rafts mediate the synaptic localization of alpha-synuclein. J Neurosci 2004, 24:6715-6723.
• [7]Nemani VM, Lu W, Berge V, Nakamura K, Onoa B, Lee MK, Chaudhry FA, Nicoll RA, Edwards RH: Increased expression of alpha-synuclein reduces neurotransmitter release by inhibiting synaptic vesicle reclustering after endocytosis. Neuron 2010, 65:66-79.
• [8]Chartier-Harlin MC, Kachergus J, Roumier C, Mouroux V, Douay X, Lincoln S, Levecque C, Larvor L, Andrieux J, Hulihan M, et al.: Alpha-synuclein locus duplication as a cause of familial Parkinson’s disease. Lancet 2004, 364:1167-1169.
• [9]Fuchs J, Nilsson C, Kachergus J, Munz M, Larsson EM, Schule B, Langston JW, Middleton FA, Ross OA, Hulihan M, et al.: Phenotypic variation in a large Swedish pedigree due to SNCA duplication and triplication. Neurology 2007, 68:916-922.
• [10]Kruger R, Kuhn W, Muller T, Woitalla D, Graeber M, Kosel S, Przuntek H, Epplen JT, Schols L, Riess O: Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson’s disease. Nat Genet 1998, 18:106-108.
• [11]Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, Pike B, Root H, Rubenstein J, Boyer R, et al.: Mutation in the alpha-synuclein gene identified in families with Parkinson’s disease. Science 1997, 276:2045-2047.
• [12]Zarranz JJ, Alegre J, Gomez-Esteban JC, Lezcano E, Ros R, Ampuero I, Vidal L, Hoenicka J, Rodriguez O, Atares B, et al.: The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia. Ann Neurol 2004, 55:164-173.
• [13]Simon-Sanchez J, Schulte C, Bras JM, Sharma M, Gibbs JR, Berg D, Paisan-Ruiz C, Lichtner P, Scholz SW, Hernandez DG, et al.: Genome-wide association study reveals genetic risk underlying Parkinson’s disease. Nat Genet 2009, 41:1308-1312.
• [14]Block ML, Hong JS: Microglia and inflammation-mediated neurodegeneration: multiple triggers with a common mechanism. Prog Neurobiol 2005, 76:77-98.
• [15]McGeer PL, Itagaki S, Akiyama H, McGeer EG: Rate of cell death in parkinsonism indicates active neuropathological process. Ann Neurol 1988, 24:574-576.
• [16]Kurkowska-Jastrzebska I, Wronska A, Kohutnicka M, Czlonkowski A, Czlonkowska A: The inflammatory reaction following 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine intoxication in mouse. Exp Neurol 1999, 156:50-61.
• [17]Benner EJ, Banerjee R, Reynolds AD, Sherman S, Pisarev VM, Tsiperson V, Nemachek C, Ciborowski P, Przedborski S, Mosley RL, Gendelman HE: Nitrated alpha-synuclein immunity accelerates degeneration of nigral dopaminergic neurons. PLoS One 2008, 3:e1376.
• [18]Ouchi Y, Yoshikawa E, Sekine Y, Futatsubashi M, Kanno T, Ogusu T, Torizuka T: Microglial activation and dopamine terminal loss in early Parkinson’s disease. Ann Neurol 2005, 57:168-175.
• [19]Hirsch EC, Hunot S: Neuroinflammation in Parkinson’s disease: a target for neuroprotection? Lancet Neurol 2009, 8:382-397.
• [20]Hunot S, Boissiere F, Faucheux B, Brugg B, Mouatt-Prigent A, Agid Y, Hirsch EC: Nitric oxide synthase and neuronal vulnerability in Parkinson’s disease. Neuroscience 1996, 72:355-363.
• [21]Hayden MS, Ghosh S: Signaling to NF-kappaB. Genes Dev 2004, 18:2195-2224.
• [22]Bonizzi G, Karin M: The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol 2004, 25:280-288.
• [23]Pasparakis M, Luedde T, Schmidt-Supprian M: Dissection of the NF-kappaB signalling cascade in transgenic and knockout mice. Cell Death Differ 2006, 13:861-872.
• [24]Ghosh A, Roy A, Liu X, Kordower JH, Mufson EJ, Hartley DM, Ghosh S, Mosley RL, Gendelman HE, Pahan K: Selective inhibition of NF-kappaB activation prevents dopaminergic neuronal loss in a mouse model of Parkinson’s disease. Proc Natl Acad Sci U S A 2007, 104:18754-18759.
• [25]Kaltschmidt B, Uherek M, Volk B, Baeuerle PA, Kaltschmidt C: Transcription factor NF-kappaB is activated in primary neurons by amyloid beta peptides and in neurons surrounding early plaques from patients with Alzheimer disease. Proc Natl Acad Sci U S A 1997, 94:2642-2647.
• [26]Kawamoto EM, Lepsch LB, Boaventura MF, Munhoz CD, Lima LS, Yshii LM, Avellar MC, Curi R, Mattson MP, Scavone C: Amyloid beta-peptide activates nuclear factor-kappaB through an N-methyl-D-aspartate signaling pathway in cultured cerebellar cells. J Neurosci Res 2008, 86:845-860.
• [27]Camandola S, Mattson MP: NF-kappa B as a therapeutic target in neurodegenerative diseases. Expert Opin Ther Targets 2007, 11:123-132.
• [28]Tiscornia G, Singer O, Verma IM: Production and purification of lentiviral vectors. Nat Protoc 2006, 1:241-245.
• [29]Rath M, Fohr KJ, Weigt HU, Gauss A, Engele J, Georgieff M, Koster S, Adolph O: Etomidate reduces glutamate uptake in rat cultured glial cells: involvement of PKA. Br J Pharmacol 2008, 155:925-933.
• [30]Larsen KE, Fon EA, Hastings TG, Edwards RH, Sulzer D: Methamphetamine-induced degeneration of dopaminergic neurons involves autophagy and upregulation of dopamine synthesis. J Neurosci 2002, 22:8951-8960.
• [31]Koh JY, Choi DW: Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Methods 1987, 20:83-90.
• [32]Lepsch LB, Munhoz CD, Kawamoto EM, Yshii LM, Lima LS, Curi-Boaventura MF, Salgado TM, Curi R, Planeta CS, Scavone C: Cocaine induces cell death and activates the transcription nuclear factor kappa-B in PC12 cells. Mol Brain 2009, 2:3. BioMed Central Full Text
• [33]Kopp E, Ghosh S: Inhibition of NF-kappa B by sodium salicylate and aspirin. Science 1994, 265:956-959.
• [34]Grilli M, Pizzi M, Memo M, Spano P: Neuroprotection by aspirin and sodium salicylate through blockade of NF-kappaB activation. Science 1996, 274:1383-1385.
• [35]Cai T, Yao T, Zheng G, Chen Y, Du K, Cao Y, Shen X, Chen J, Luo W: Manganese induces the overexpression of alpha-synuclein in PC12 cells via ERK activation. Brain Res 2010, 1359:201-207.
• [36]Beraud D, Hathaway HA, Trecki J, Chasovskikh S, Johnson DA, Johnson JA, Federoff HJ, Shimoji M, Mhyre TR, Maguire-Zeiss KA: Microglial activation and antioxidant responses induced by the Parkinson’s disease protein alpha-synuclein. J Neuroimmune Pharmacol 2013, 8:94-117.
• [37]Banati RB, Daniel SE, Blunt SB: Glial pathology but absence of apoptotic nigral neurons in long-standing Parkinson’s disease. Mov Disord 1998, 13:221-227.
• [38]Nagatsu T, Mogi M, Ichinose H, Togari A. Changes in cytokines and neurotrophins in Parkinson’s disease. J Neural Transm Suppl. 2000;(60):277-290
• [39]Nagai M, Re DB, Nagata T, Chalazonitis A, Jessell TM, Wichterle H, Przedborski S: Astrocytes expressing ALS-linked mutated SOD1 release factors selectively toxic to motor neurons. Nat Neurosci 2007, 10:615-622.
• [40]Di Giorgio FP, Carrasco MA, Siao MC, Maniatis T, Eggan K: Non-cell autonomous effect of glia on motor neurons in an embryonic stem cell-based ALS model. Nat Neurosci 2007, 10:608-614.
• [41]Gentle IE, Wong WW, Evans JM, Bankovacki A, Cook WD, Khan NR, Nachbur U, Rickard J, Anderton H, Moulin M, et al.: In TNF-stimulated cells, RIPK1 promotes cell survival by stabilizing TRAF2 and cIAP1, which limits induction of non-canonical NF-kappaB and activation of caspase-8. J Biol Chem 2011, 286:13282-13291.
• [42]Varfolomeev E, Goncharov T, Maecker H, Zobel K, Komuves LG, Deshayes K, Vucic D: Cellular inhibitors of apoptosis are global regulators of NF-kappaB and MAPK activation by members of the TNF family of receptors. Sci Signal 2012, 5:ra22.
• [43]Lazaro DF, Rodrigues EF, Langohr R, Shahpasandzadeh H, Ribeiro T, Guerreiro P, Gerhardt E, Krohnert K, Klucken J, Pereira MD, et al.: Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation. PLoS Genet 2014, 10:e1004741.
• [44]Beg AA, Baltimore D: An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science 1996, 274:782-784.
• [45]Rojanathammanee L, Murphy EJ, Combs CK: Expression of mutant alpha-synuclein modulates microglial phenotype in vitro. J Neuroinflammation 2011, 8:44. BioMed Central Full Text
• [46]Taha TA, Kitatani K, Bielawski J, Cho W, Hannun YA, Obeid LM: Tumor necrosis factor induces the loss of sphingosine kinase-1 by a cathepsin B-dependent mechanism. J Biol Chem 2005, 280:17196-17202.
• [47]Sarnico I, Lanzillotta A, Boroni F, Benarese M, Alghisi M, Schwaninger M, Inta I, Battistin L, Spano P, Pizzi M: NF-kappaB p50/RelA and c-Rel-containing dimers: opposite regulators of neuron vulnerability to ischaemia. J Neurochem 2009, 108:475-485.