Locomotor and histological changes in a cuprizone-induced animal model of multiple sclerosis: comparison between alpha-tocopherol and fingolimod

Nilesh Kumar Mitra , Nermesh Singh A/L Gurdib Singh, Nurul Ain Najihah Binti Wadingasafi, Jestin Chellian


Background and purpose: Fingolimod is a sphingosine 1-phosphate receptor modulator used to treat multiple sclerosis (MS). Alpha-tocopherol (AT) has been found to improve motor function in an animal model of MS. In the present study, the effects of AT and fingolimod on the locomotor function and histological evidence of demyelination were compared in a cuprizone-induced rat model of MS.

Experimental approach: Female Sprague-Dawley rats (8 weeks) were fed with 0.2% (w/w) cuprizone diet for 5 weeks followed by intraperitoneal injections of fingolimod (3 mg/Kg; group F, n = 10) and alpha-tocopherol (100 mg/Kg; group A, n = 10). Vehicle-treated rats (group V, n = 10) were treated intraperitoneally with 1% ethanol in saline on weeks 6 and 7. Open field and beam walking tests were carried out every 10 days. The mean area of demyelination in the corpus callosum was quantified using Luxol fast blue stained histological sections of the forebrain.

Findings/Results: The mean speed of movement was increased by 54% and 50% in groups F and A compared to group V. Total distance moved was increased by 61% and 52.7% in groups F and A compared to group V. Mean time to walk the beam was reduced in group A by 52% compared to group V. Mean frequency of crossing lines from the inner squares to outer squares was reduced in groups A and F compared to group V. Mean area of demyelination in corpus callosum showed 62% reduction in group A compared to group V.

Conclusion and implications: Both fingolimod and AT treatments improved the locomotor function. However, AT treatment reduced the areas of demyelination in higher proportion and improved motor coordination and exploratory behavior.




Keywords: Alpha-tocopherol; Demyelination; Fingolimod; Cuprizone; Multiple sclerosis.

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Kipp M. Remyelination strategies in multiple sclerosis: a critical reflection. Expert Rev Neurother. 2016;16(1):1-3.

DOI: 10.1586/14737175.2016.1116387.

Torkildsen O, Brunborg LA, Myhr KM, Bø L. The cuprizone model for demyelination. Acta Neurol Scand Suppl. 2008;188:72-76.

DOI: 10.1111/j.1600-0404.2008.01036.x.

Zendedel A, Beyer C, Kipp M. Cuprizone-induced demyelination as a tool to study remyelination and axonal protection. J Mol Neurosci. 2013;51(2):567-572.

DOI: 10.1007/s12031-013-0026-4.

Chastain EM, Duncan DS, Rodgers JM, Miller SD. The role of antigen presenting cells in multiple sclerosis. Biochim Biophys Acta. 2011;1812(2): 265-274.

DOI: 10.1016/j.bbadis.2010.07.008.

Richards RG, Sampson FC, Beard SM, Tappenden P. A review of the natural history and epidemiology of multiple sclerosis: implications for resource allocation and health economic models. Health Technol Assess. 2002;6(10):1-73.

DOI: 10.3310/hta6100.

Rosen H, Gonzalez-Cabrera PJ, Sanna MG, Brown S. Sphingosine 1-phosphate receptor signaling. Annu Rev Biochem. 2009;78:743-768.

DOI: 10.1146/annurev.biochem.78.072407.103733.

Hunter SF, Bowen JD, Reder AT. The direct effects of fingolimod in the central nervous system: implications for relapsing multiple sclerosis. CNS Drugs. 2016;30(2):135-147.

DOI: 10.1007/s40263-015-0297-0.

EDQM, Council of Europe. The European Pharmacopeia. 5th ed. Supplement 5.1. Strasbourg, France: EDQM Council of Europe; 2005. pp:3024.

Ferri P, Angelino D, Gennari L, Benedetti S, Ambrogini P, Del Grande, et al. Enhancement of flavonoid ability to cross the blood-brain barrier of rats by co-administration with α-tocopherol. Food Funct. 2015;6(2):394-400.

DOI: 10.1039/c4fo00817k.

Seibenhener ML, Wooten MC. Use of the open field maze to measure locomotor and anxiety-like behavior in mice. J Vis Exp. 2015;96:e52434,1-6.

DOI: 10.3791/52434.

Luong TN, Carlisle HJ, Southwell A, Patterson PH. Assessment of motor balance and coordination in mice using the balance beam. J Vis Exp. 2011;49:2376,1-3.

DOI: 10.3791/2376.

Mitra NK, Xuan KY, Teo CC, Xian-Zhuang N, Singh A, Chellian J. Evaluation of neuroprotective effects of alpha-tocopherol in cuprizone-induced demyelination model of multiple sclerosis. Res Pharm Sci. 2020;15(6):602-611.

DOI: 10.4103/1735-5362.301345.

Xue H, Ren H, Zhang L, Sun X, Wang W, Zhang S, et al. Alpha-tocopherol ameliorates experimental autoimmune encephalomyelitis through the regulation of Th1 cells. Iran J Basic Med Sci. 2016;19(5):561-566.

PMID: 27403263.

Li L, Matsumoto M, Seabrook TJ, Cojean C, Brinkman V, Pachner AR. The effect of FTY720 in the Theiler's virus model of multiple sclerosis. J Neurol Sci. 2011;308(1-2):41-48.

DOI: 10.1016/j.jns.2011.06.029.

Mao Y, Wang J, Zhao Y, Wu Y, Kwak KJ, Chen CS, et al. A novel liposomal formulation of FTY720 (fingolimod) for promising enhanced targeted delivery. Nanomedicine. 2014;10(2):393-400.

DOI: 10.1016/j.nano.2013.08.001.

Igado OO, Andrioli A, Azeez IA, Girolamo F, Errede M, Aina OO, et al. The ameliorative effects of a phenolic derivative of Moringa oleifera leave against vanadium-induced neurotoxicity in mice. IBRO Rep. 2020;9:164-182.

DOI: 10.1016/j.ibror.2020.07.004.

Masood A, Banerjee B, Vijayan VK, Ray A. Modulation of stress-induced neurobehavioral changes by nitric oxide in rats. Eur J Pharmacol. 2003;458(1-2):135-139.

DOI: 10.1016/s0014-2999(02)02688-2.

Ramis MR, Sarubbo F, Terrasa JL, Moranta D, Aparicio S, Miralles A, et al. Chronic α-tocopherol increases central monoamines synthesis and improves cognitive and motor abilities in old rats. Rejuvenation Res. 2016;19(2):159-171.

DOI: 10.1089/rej.2015.1685.

Sestakova N, Puzserova A, Kluknavsky M, Bernatova I. Determination of motor activity and anxiety-related behaviour in rodents: methodological aspects and role of nitric oxide. Interdiscip Toxicol. 2013;6(3):126-135.

DOI: 10.2478/intox-2013-0020.

Mitra NK, Nadarajah VD, Siong HH. Effect of concurrent application of heat, swim stress and repeated dermal application of chlorpyrifos on the hippocampal neurons in mice. Folia Neuropathol. 2009;47(1):60-68.

PMID: 19353435.

Mohammadi-Rad M, Ghasemi N, Aliomrani M. Evaluation of apamin effects on myelination process in C57BL/6 mice model of multiple sclerosis. Res Pharm Sci. 2019;14(5):424-431.

DOI: 10.4103/1735-5362.268203.

Procaccini C, De Rosa V, Pucino V, Formisano L, Matarese G. Animal models of multiple sclerosis. Eur J Pharmacol. 2015;759:182-191.

DOI: 10.1016/j.ejphar.2015.03.042.

Pepper RE, Pitman KA, Cullen CL, Young KM. How do cells of the oligodendrocyte lineage affect neuronal circuits to influence motor function, memory and mood? Front Cell Neurosci. 2018;12:399-412.

DOI: 10.3389/fncel.2018.00399.

Jung CG, Kim HJ, Miron VE, Cook S, Kennedy TE, Foster CA, et al. Functional consequences of S1P receptor modulation in rat oligodendroglial lineage cells. Glia. 2007;55(16):1656-1667.

DOI: 10.1002/glia.20576.

Bonfiglio T, Olivero G, Merega E, Di Prisco S, Padolecchia C, Grilli M, et al. Prophylactic versus therapeutic fingolimod: restoration of presynaptic defects in mice suffering from experimental autoimmune encephalomyelitis. PLoS One. 2017;12(1):e0170825,1-29.

DOI: 10.1371/journal.pone.0170825.

Landi D, Vollaro S, Pellegrino G, Mulas D, Ghazaryan A, Falato E, et al. Oral fingolimod reduces glutamate-mediated intracortical excitability in relapsing-remitting multiple sclerosis. Clin Neurophysiol. 2015;126(1):165-169.

DOI: 10.1016/j.clinph.2014.05.031.

de Carvalho TS, Cardoso PB, Santos-Silva M, Lima-Bastos S, Luz WL, Assad N, et al. Oxidative stress mediates anxiety-like behavior induced by high caffeine intake in zebrafish: protective effect of alpha-tocopherol. Oxid Med Cell Longev. 2019;2019:8419810,1-9.

DOI: 10.1155/2019/8419810.

Barth AM, Domonkos A, Fernandez-Ruiz A, Freund TF, Varga V. Hippocampal network dynamics during rearing episodes. Cell Rep. 2018;23(6):1706-1715.

DOI: 10.1016/j.celrep.2018.04.021.

Ambrogini P, Betti M, Galati C, Di Palma M, Lattanzi D, Savelli D, et al. α-Tocopherol and hippocampal neural plasticity in physiological and pathological conditions. Int J Mol Sci. 2016;17(12):2107-2138.

DOI: 10.3390/ijms17122107.

Hu Y, Lee X, Ji B, Guckian K, Apicco D, Pepinsky R, et al. Sphingosine 1-phosphate receptor modulator fingolimod (FTY720) does not promote remyelination in vivo. Mol Cell Neurosci. 2011;48(1):72-81.

DOI: 10.1016/j.mcn.2011.06.007.

Slowik A, Schmidt T, Beyer C, Amor S, Clarner T, Kipp M. The sphingosine 1-phosphate receptor agonist FTY720 is neuroprotective after cuprizone-induced CNS demyelination. Br J Pharmacol. 2015;172(1):80-92.

DOI: 10.1111/bph.12938.

Blanchard B, Heurtaux T, Garcia C, Moll NM, Caillava C, Grandbarbe L, et al. Tocopherol derivative TFA-12 promotes myelin repair in experimental models of multiple sclerosis. J Neurosci. 2013;33(28):11633-11642.

DOI: 10.1523/JNEUROSCI.0774-13.2013.

Franklin RJ. Why does remyelination fail in multiple sclerosis? Nat Rev Neurosci. 2002;3(9):705-714.

DOI: 10.1038/nrn917.

O'Sullivan S, Dev KK. Sphingosine-1-phosphate receptor therapies: advances in clinical trials for CNS-related diseases. Neuropharmacology. 2017;113(Pt B):597-607.

DOI: 10.1016/j.neuropharm.2016.11.006.

Yazdi A, Ghasemi-Kasman M, Javan M. Possible regenerative effects of fingolimod (FTY720) in multiple sclerosis disease: an overview on remyelination process. J Neurosci Res. 2020;98(3):524-536.

DOI: 10.1002/jnr.24509.


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