Neuroprotective effect of chondroitin sulfate on SH‑SY5Y cells overexpressing wild‑type or A53T mutant α‑synuclein

Ju,Chuanxia; Gao,Jianjun; Hou,Lin; Wang,Lei; Zhang,Fang; Sun,Fusheng; Zhang,Tingting; Xu,Pingping; Shi,Zhenyan; Hu,Fang; Zhang,Congxiao

doi:10.3892/mmr.2017.7725

Neuroprotective effect of chondroitin sulfate on SH‑SY5Y cells overexpressing wild‑type or A53T mutant α‑synuclein

Authors:
- Chuanxia Ju
- Jianjun Gao
- Lin Hou
- Lei Wang
- Fang Zhang
- Fusheng Sun
- Tingting Zhang
- Pingping Xu
- Zhenyan Shi
- Fang Hu
- Congxiao Zhang
View Affiliations

Affiliations: Department of Pharmacology, Qingdao University School of Pharmacy, Qingdao, Shandong 266021, P.R. China, School of Basic Medical Sciences, Qingdao University, Qingdao, Shandong 266021, P.R. China, Qingdao Municipal Hospital, Qingdao, Shandong 266011, P.R. China, College of Marine Life Sciences, Ocean University of China, Qingdao, Shandong 266003, P.R. China
Published online on: October 4, 2017 https://doi.org/10.3892/mmr.2017.7725
Pages: 8721-8728
Copyright: © Ju et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Accumulation of α‑synuclein (α‑SYN) is a common pathology for Parkinson's disease (PD). There is abundant evidence that the toxic‑gain‑of‑function of α‑SYN's is associated with aggregation and consequent effects. To assess the potential of chondroitin sulfate (CS) in this regard, the present study investigated its neuroprotective on SH‑SY5Y cells overexpressing wild‑type (WT) or A53T mutant α‑SYN. Cell viability was measured by MTT assay. Apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential were detected by flow cytometry. The protein expression levels of total α‑SYN, phosphorylated Ser129 α‑SYN, B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein (Bax) and cytochrome‑c (Cyt‑c ) were analyzed by western blotting. It was observed that CS reduced the expression levels of total α‑SYN and phosphorylated Ser129 α‑SYN, prevented cell loss and inhibited apoptosis. The subsequent mechanism study indicated that CS inhibited ROS overproduction. CS also significantly attenuated WT and A53T mutant α‑SYN‑induced dysfunction, including decrease of mitochondrial membrane potential, decrease of Bcl‑2 expression, and increase of Bax expression, release of Cyt‑c from the mitochondria and activation of caspase‑3 and caspase‑9, which demonstrated that CS suppressed α‑SYN‑induced apoptosis possibly through mitochondria protection. These results suggested that CS protects SH‑SY5Y cells overexpressing WT or A53T mutant α‑SYN by inhibiting the expression and phosphorylation of α‑SYN, and ROS overproduction and mitochondrial apoptosis. These results implicate CS as a potential therapeutic agent for the treatment of PD.

View Figures

View References

1	Chiba-Falek O, Lopez GJ and Nussbaum RL: Levels of alpha-synuclein mRNA in sporadic Parkinson disease patients. Mov Disord. 21:1703–1708. 2006. View Article : Google Scholar : PubMed/NCBI
2	Singleton AB, Farrer M, Johnson J, Singleton A, Hague S, Kachergus J, Hulihan M, Peuralinna T, Dutra A, Nussbaum R, et al: alpha-Synuclein locus triplication causes Parkinson's disease. Science. 302:8412002. View Article : Google Scholar
3	Galtrey CM and Fawcett JW: The role of chondroitin sulfate proteoglycans in regeneration and plasticity in the central nervous system. Brain Res Rev. 54:1–18. 2007. View Article : Google Scholar : PubMed/NCBI
4	Purushothaman A, Fukuda J, Mizumoto S, ten Dam GB, van Kuppevelt TH, Kitagawa H, Mikami T and Sugahara K: Functions of chondroitin sulfate/dermatan sulfate chains in brain development. Critical roles of E and iE disaccharide units recognized by a single chain antibody GD3G7. J Biol Chem. 282:19442–19452. 2007. View Article : Google Scholar : PubMed/NCBI
5	Okamoto M, Mori S, Ichimura M and Endo H: Chondroitin sulfate proteoglycans protect cultured rat's cortical and hippocampal neurons from delayed cell death induced by excitatory amino acids. Neurosci Lett. 172:51–54. 1994. View Article : Google Scholar : PubMed/NCBI
6	Sato Y, Nakanishi K, Tokita Y, Kakizawa H, Ida M, Maeda H, Matsui F, Aono S, Saito A, Kuroda Y, et al: A highly sulfated chondroitin sulfate preparation, CS-E, prevents excitatory amino acid-induced neuronal cell death. J Neurochem. 104:1565–1576. 2008. View Article : Google Scholar : PubMed/NCBI
7	Cañas N, Valero T, Villarroya M, Montell E, Vergés J, García AG and López MG: Chondroitin sulfate protects SH-SY5Y cells from oxidative stress by inducing heme oxygenase-1 via phosphatidylinositol 3-kinase/Akt. J Pharmacol Exp Ther. 323:946–953. 2007. View Article : Google Scholar : PubMed/NCBI
8	Ju C, Hou L, Sun F, Zhang L, Zhang Z, Gao H, Wang L, Wang D, Lv Y and Zhao X: Anti-oxidation and antiapoptotic effects of chondroitin sulfate on 6-hydroxydopamine-induced injury through the up-regulation of nrf2 and inhibition of mitochondria-mediated pathway. Neurochem Res. 40:1509–1519. 2015. View Article : Google Scholar : PubMed/NCBI
9	McLaughlin RW, De Stigter JK, Sikkink LA, Baden EM and Ramirez-Alvarado M: The effects of sodium sulfate, glycosaminoglycans, and Congo red on the structure, stability, and amyloid formation of an immunoglobulin light-chain protein. Protein Sci. 15:1710–1722. 2006. View Article : Google Scholar : PubMed/NCBI
10	Zhang Q, Li J, Liu C, Song C, Li P, Yin F, Xiao Y, Li J, Jiang W, Zong A, et al: Protective effects of low molecular weight chondroitin sulfate on amyloid beta (Aβ)-induced damage in vitro and in vivo. Neuroscience. 305:169–182. 2015. View Article : Google Scholar : PubMed/NCBI
11	Woods AG, Cribbs DH, Whittemore ER and Cotman CW: Heparan sulfate and chondroitin sulfate glycosaminoglycan attenuate beta-amyloid(25–35) induced neurodegeneration in cultured hippocampal neurons. Brain Res. 697:53–62. 1995. View Article : Google Scholar : PubMed/NCBI
12	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. 364:1167–1169. 2004. View Article : Google Scholar : PubMed/NCBI
13	Paumier KL, Rizzo SJ Sukoff, Berger Z, Chen Y, Gonzales C, Kaftan E, Li L, Lotarski S, Monaghan M, Shen W, et al: Behavioral characterization of A53T mice reveals early and late stage deficits related to Parkinson's disease. PLoS One. 8:e702742013. View Article : Google Scholar : PubMed/NCBI
14	Chesselet MF: In vivo alpha-synuclein overexpression in rodents: A useful model of Parkinson's disease? Exp Neurol. 209:22–27. 2008. View Article : Google Scholar : PubMed/NCBI
15	Lashuel HA and Hirling H: Rescuing defective vesicular trafficking protects against alpha-synuclein toxicity in cellular and animal models of Parkinson's disease. ACS Chem Biol. 1:420–424. 2006. View Article : Google Scholar : PubMed/NCBI
16	Periquet M, Fulga T, Myllykangas L, Schlossmacher MG and Feany MB: Aggregated alpha-synuclein mediates dopaminergic neurotoxicity in vivo. J Neurosci. 27:3338–3346. 2007. View Article : Google Scholar : PubMed/NCBI
17	Saha AR, Ninkina NN, Hanger DP, Anderton BH, Davies AM and Buchman VL: Induction of neuronal death by alpha-synuclein. Eur J Neurosci. 12:3073–3077. 2000. View Article : Google Scholar : PubMed/NCBI
18	Hsu LJ, Sagara Y, Arroyo A, Rockenstein E, Sisk A, Mallory M, Wong J, Takenouchi T, Hashimoto M and Masliah E: alpha-synuclein promotes mitochondrial deficit and oxidative stress. Am J Pathol. 157:401–410. 2000. View Article : Google Scholar : PubMed/NCBI
19	Parihar MS, Parihar A, Fujita M, Hashimoto M and Ghafourifar P: Mitochondrial association of alpha-synuclein causes oxidative stress. Cell Mol Life Sci. 65:1272–1284. 2008. View Article : Google Scholar : PubMed/NCBI
20	Parihar MS, Parihar A, Fujita M, Hashimoto M and Ghafourifar P: Alpha-synuclein overexpression and aggregation exacerbates impairment of mitochondrial functions by augmenting oxidative stress in human neuroblastoma cells. Int J Biochem Cell Biol. 41:2015–2024. 2009. View Article : Google Scholar : PubMed/NCBI
21	Martin LJ, Semenkow S, Hanaford A and Wong M: Mitochondrial permeability transition pore regulates Parkinson's disease development in mutant α-synuclein transgenic mice. Neurobiol Aging. 35:1132–1152. 2014. View Article : Google Scholar : PubMed/NCBI
22	Anderson JP, Walker DE, Goldstein JM, de Laat R, Banducci K, Caccavello RJ, Barbour R, Huang J, Kling K, Lee M, et al: Phosphorylation of Ser-129 is the dominant pathological modification of alpha-synuclein in familial and sporadic Lewy body disease. J Biol Chem. 281:29739–29752. 2006. View Article : Google Scholar : PubMed/NCBI
23	Fujiwara H, Hasegawa M, Dohmae N, Kawashima A, Masliah E, Goldberg MS, Shen J, Takio K and Iwatsubo T: alpha-Synuclein is phosphorylated in synucleinopathy lesions. Nat Cell Biol. 4:160–164. 2002.PubMed/NCBI
24	Sugeno N, Takeda A, Hasegawa T, Kobayashi M, Kikuchi A, Mori F, Wakabayashi K and Itoyama Y: Serine 129 phosphorylation of alpha-synuclein induces unfolded protein response-mediated cell death. Biol Chem. 283:23179–23188. 2008. View Article : Google Scholar
25	Sato H, Arawaka S, Hara S, Fukushima S, Koga K, Koyama S and Kato T: Authentically phosphorylated α-synuclein at Ser129 accelerates neurodegeneration in a rat model of familial Parkinson's disease. J Neurosci. 31:16884–16894. 2011. View Article : Google Scholar : PubMed/NCBI
26	Oliveras-Salvá M, Van der Perren A, Casadei N, Stroobants S, Nuber S, D'Hooge R, Van den Haute C and Baekelandt V: rAAV2/7 vector-mediated overexpression of alpha-synuclein in mouse substantia nigra induces protein aggregation and progressive dose-dependent neurodegeneration. Mol Neurodegener. 8:442013. View Article : Google Scholar : PubMed/NCBI
27	Dias V, Junn E and Mouradian MM: The role of oxidative stress in Parkinson's disease. J Parkinsons Dis. 3:461–491. 2013.PubMed/NCBI
28	Junn E and Mouradian MM: Human alpha-synuclein over-expression increases intracellular reactive oxygen species levels and susceptibility to dopamine. Neurosci Lett. 320:146–150. 2002. View Article : Google Scholar : PubMed/NCBI
29	Devi L, Raghavendran V, Prabhu BM, Avadhani NG and Anandatheerthavarada HK: Mitochondrial import and accumulation of alpha-synuclein impair complex I in human dopaminergic neuronal cultures and Parkinson disease brain. J Biol Chem. 283:9089–9100. 2008. View Article : Google Scholar : PubMed/NCBI
30	Chinta SJ, Mallajosyula JK, Rane A and Andersen JK: Mitochondrial α-synuclein accumulation impairs complex I function in dopaminergic neurons and results in increased mitophagy in vivo. Neurosci Lett. 486:235–239. 2010. View Article : Google Scholar : PubMed/NCBI
31	Zhu Y, Duan C, Lü L, Gao H, Zhao C, Yu S, Uéda K, Chan P and Yang H: α-Synuclein overexpression impairs mitochondrial function by associating with adenylate translocator. Int J Biochem Cell Biol. 43:732–741. 2011. View Article : Google Scholar : PubMed/NCBI
32	Seo JH, Rah JC, Choi SH, Shin JK, Min K, Kim HS, Park CH, Kim S, Kim EM, Lee SH, et al: α-synuclein regulates neuronal survival via Bcl-2 family expression and PI3/Akt kinase pathway. FASEB J. 16:1826–1828. 2002.PubMed/NCBI
33	Yuan Y, Jin J, Yang B, Zhang W, Hu J, Zhang Y and Chen NH: Overexpressed alpha-synuclein regulated the nuclear factor-kappaB signal pathway. Cell Mol Neurobiol. 28:21–33. 2008. View Article : Google Scholar : PubMed/NCBI
34	Niture SK, Kaspar JW, Shen J and Jaiswal AK: Nrf2 signaling and cell survival. Toxicol Appl Pharmacol. 244:37–42. 2010. View Article : Google Scholar : PubMed/NCBI
35	Hwang YP and Jeong HG: Mechanism of phytoestrogen puerarin-mediated cytoprotection following oxidative injury: Estrogen receptor-dependent up-regulation of PI3K/Akt and HO-1. Toxicol Appl Pharmacol. 233:371–381. 2008. View Article : Google Scholar : PubMed/NCBI