Abstract
Impaired mitochondrial function in glial and neuronal cells in the substantia nigra is one of the most likely causes of Parkinson’s disease. In this study, we investigated the protective role of glucose on early key events associated with MPP+-induced changes in rat C6 astroglial cells. Studies were carried out to examine alterations in mitochondrial respiratory status, membrane potential, glutathione levels, and cell cycle phase inhibition at 48 h in 2 and 10 mM glucose in media. The results obtained suggest that MPP+ caused significant cell death in 2 mM glucose with LC50 0.14 ± 0.005 mM, while 10 mM glucose showed highly significant protection against MPP+ toxicity with LC50 0.835 ± 0.03 mM. This protection was not observed with cocaine, demonstrating its compound specificity. MPP+ in 2 mM glucose decreased significantly mitochondrial respiration, membrane potential and glutathione levels in a dose dependent manner, while 10 mM glucose significantly restored them. MPP+ in 2 mM glucose arrested the cells at G0/G1 and G2/M phases, demonstrating its dual inhibitory effects. However, in 10 mM glucose, MPP+ caused G0/G1 arrest only. In summary, the results suggest that loss of cell viability in 2 mM glucose group with MPP+ treatments was due to mitochondrial dysfunction caused by multilevel mechanism, involving significant decrease in mitochondrial respiration, membrane potential, glutathione levels, and dual arrest of cell phases, while 10 mM glucose rescued astroglial cells from MPP+ toxicity by significant maintenance of these factors.
Similar content being viewed by others
References
Langston JW, Langston EB, Irwin I (1984) MPTP-induced parkinsonism in human and non-human primates: clinical and experimental aspects. Acta Neurol Scand 100:49–54 (Suppl.)
Hunot F, Boissiere F, Faucheux B (1996) Nitric oxide synthase and neuronal vulnerability in Parkinson’s disease. Neuroscience 72:355–363
McGeer PL, Itagaki S, Boyes BE, McGeer EG (1988) Reactive microglia are positive for HLA-DR in the substantia nigra of Parkinson’s and Alzheimer’s disease brains. Neurology 38:1285–1291
Hunot B, Brugg B, Ricard D (1997) Nuclear translocation of NF-kappa B is increased in dopaminergic neurons of patients with Parkinson’ disease. Proc Natl Acad Sci USA 94:7531–7536
Blass JP (2002) Glucose/mitochondria in neurological conditions. Int Rev Neurobiol 51:325–376
Heikkila RE, Manzino L, Duvoisin RC, Cabbat FS (1984) Protection against the dopaminergic neurotoxicity of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine by monoamine oxidase inhibitors. Nature 311:467–469
Markey SP, Johannessen JN, Chiueh CC, Burns RS, Herkenham MA (1984) Intraneuronal generation of a pyridinium metabolite may cause drug-induced parkinsonism. Nature 311:464–467
Westlund KN, Denney RM, Kochersperger LM, Rose RM, Abell CW (1985) Distinct monoamine oxidase A and B populations in primate brain. Science 230:181–183
Takada M, Li ZK, Hattori T (1990) Astroglial ablation prevents MPTP-induced nigrostriatal neuronal death. Brain Res 509:55–61
Di Monte DA (1991) Mitochondrial DNA and Parkinson’s disease. Neurology 41([suppl. 2]):38–42
Forno LS, Delanney LE, Irwin I, Di Monte D, Langston JW (1992) Astrocytes and Parkinson’s disease. Prog Brain Res 94:429–436
Singer TP, Ramsay RR (1990) Mechanism of the neurotoxicity of MPTP: an update. FEBS Lett 274:1–8
Mazzio E, Soliman KFA (2003) The role of glycolysis and gluconeogenesis in the cytoprotection of neuroblastoma cells against 1-methyl-3-phenyl-pyridinium ion toxicity. Neurotoxicology 24:137–147
Itano Y, Kitamura Y, Nomura Y (1995) Biphasic effects of MPP+, a possible parkinsonism inducer, on dopamine content and tyrosine hydroxylase mRNA expression in PC12 cells. Neurochem Int 26:165–171
Sheehan JP, Palmer PE, Helm GA, Tuttle JB (1997) MPP+ induced apoptotic cell death in SH-SY5Y neuroblastoma cells: an electron microscope study. J Neurosci Res 48:226–237
De Girolamo LA, Hargreaves AJ, Billett EE (2001) Proection from MPTP-induced neurotoxicity in differentiating mouse N2a neuroblastoma cells. J Neurochem 76:650–660
Lund-Andersen H (1979) Transport of glucose from blood brain to brain. Physiol Rev 59:305–352
McGowan TA, Dunn SR, Falkner B, Sharma K (2006) Stimulation of urinary TGF-b and isoprostanes in response to hyperglycemia in humans. Clin J Am Soc Nephrol 1:263–268
Badisa RB, Tzakou O, Couladis M, Pilarinou E (2003) Cytotoxic activities of some Greek Labiatae herbs. Phytother Res 17:472–476
Badisa RB, Darling-Reed SF, Goodman CB (2010) Cocaine induces alterations in mitochondrial membrane potential and dual cell cycle arrest in rat C6 astroglial cells. Neurochem Res 35(2):288–297
Denizot R, Lang R (1986) Rapid colorimetric assay for cell growth and survival. J Immunol Methods 89:271–277
Smith IK, Vierheller TL, Thorne CA (1988) Assay of glutathione reductase in crude tissue homogenates using 5, 5’—dithiobis(2-nitrobenzoic acid). Anal Biochem 175:408–413
Badisa RB, Darling-Reed SF, Joseph P, Cooperwood JS, Latinwo LM, Goodman CB (2009) Selective cytotoxic activities of two novel synthetic drugs on human breast carcinoma MCF-7 cells. Anticancer Res 29:2993–2996
Ipsen J, Feigl P (1970) Bancroft’s introduction to biostatistics. Harper & Row, New York, p 455
Williams ZR, Goodman CB, Soliman KFA (2007) Anaerobic glycolysis protection against 1-methyl-4-phenyl pyridinium (MPP+) toxicity in C6 glial cells. Neurochem Res 32:1071–1080
Rollema H, Skolnik M, D’Engelbronner J, Igarashi K, Usuki E, Castagnoli N (1994) MPP (+)-like neurotoxicity of a pyridinium metabolite derived from haloperidol: in vivo microdialysis and in vitro mitochondrial studies. J Pharmacol Exp Ther 268:380–387
Cunha-Oliveira T, Rego AC, Cardoso SM, Borges F, Swerdlow RH, Macedo T, de Oliveira CR (2006) Mitochondrial dysfunction and caspase activation in rat cortical neurons treated with cocaine or amphetamine. Brain Res 1089:44–54
Lambert CE, Bondy SC (1989) Effects of MPTP, MPP+ and paraquat on mitochondrial potential and oxidative stress. Life Sci 44:1277–1284
Chalmers-Redman RME, MacLean Fraser AD, Carlile GW, Pong A, Tatton WG (1999) Glucose protection from MPP+—induced apoptosis depends on mitochondrial membrane potential and ATP synthase. Biochem Biophys Res Commun 257:440–447
Bai J, Nakamura H, Ueda S, Kwon YW, Tanaka T, Ban S, Yodoi J (2004) Proteasome-dependent degradation of cyclin D1 in 1-methyl-4-phenylpyridinium ion (MPP+)-induced cell cycle arrest. J Biol Chem 279:38710–38714
Basma AN, Heikkila RE, Saporito MS, Philbert M, Geller HM, Nicklas WJ (1992) 1-methyl-4-(2’-ethylphenyl)-1, 2, 3, 6-tetrahydropyridine-inducedtoxicity in PC12 cells is enhanced by preventing glycolysis. J Neurochem 58:1052–1059
Chan P, Langston JW, Irwin I, Delanney LE, Di Monte DA (1993) 2-deoxyglucose enhances 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine-induced ATP loss in the mouse brain. J Neurochem 61:610–616
Mizuno Y, Saitoh T, Sone N (1987) Inhibition of mitochondrial NADH-ubiquinone oxidoreductase activity by 1-methylphenylpyridinium ion. Biochem Biophys Res Commun 143:294–299
Scotcher KP, Irwin I, Delanney LE, Langston JW, Di Monte D (1990) Effects of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine and 1-methyl-4-phenylpyridinium ion on ATP levels of mouse synaptosomes. J Neurochem 54:1295–1301
Lotharius J, Dugan LL, O’Malley KL (1999) Distinct mechanisms underlie neurotoxin-mediated cell death in cultured dopaminergic neurons. J Neurosci 19:1284–1293
Amao Y, Miyashita T, Okura I (2001) Novel optical oxygen sensing material:platinum octaethylporphyrin immobilized in a copolymer film of isobutyl methacrylate and tetrafluoropropyl methacrylate. React Funct Polym 47:40–54
Mamchaoui K, Saumon G (2000) A method for measuring the oxygen consumption of intact cell monolayers. Am J Physiol Lung C 278:L858–L863
Starlinder H, Lubber DW (1972) Methodical studies on the polarographic measurement of respiration and critical oxygen pressure in mitochondria and isolated cells with membrane-covered platinum electrodes. Pflugers Archiv 337:19–28
Vanderkooi JM, Erecinska M, Silver IA (1991) Oxygen in mammalian tissue: methods of measurement and affinities of various reactions. Am J Physiol 260:1131–1150
Johnson LV, Walsh ML, Chen LB (1980) Localization of mitochondria in living cells with Rhodamine-123. Proc Natl Acad Sci USA 77:990–994
Ramsay RE, Kowal AT, Johnson MK, Salach JI, Singer TP (1987) The inhibition site of MPP+, the neurotoxic bioactivation product of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine is near the Q-binding site of NADH dehydrogenase. Arch Biochem Biophys 259:645–649
Di Monte DA, Wu EY, Delanney LE, Irwin I, Langston JW (1992) Toxicity of 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine in primary cultures of mouse astrocytes. J Pharmacol Exp Ther 261:44–49
Elsisi N, Darling-Reed S, Lee EY, Oriaku ET, Soliman KF (2005) Ibuprofen and apigenin induce apoptosis and cell cycle arrest in activated microglial. Neuroscience Lett 375:91–96
Long-Smith CM, Sullivan AM, Nolan YM (2009) The influence of microglia on the pathogenesis of Parkinson’s disease. Prog Neurobiol 89:277–287
Acknowledgments
The authors acknowledge the critical reading of the manuscript and the valuable suggestions of Dr. Sandra Suther. This study was supported by a grant obtained from the National Institutes of Health, Division of Research Resources, Research Centers in Minority Institution (RCMI) G12 RR 03020.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Badisa, R.B., Darling-Reed, S.F. & Soliman, K.F.A. The Protective Role of d-Glucose Against 1-Methyl-4-Phenylpyridinium Ion (MPP+): Induced Mitochondrial Dysfunction in C6 Astroglial Cells. Neurochem Res 35, 1413–1421 (2010). https://doi.org/10.1007/s11064-010-0200-9
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11064-010-0200-9