Comparison of the Expression Levels of MBP and GFAP Following the Induction of Neuronal Demyelination with Ethidium Bromide in Male and Female Rats

Document Type : Original Article


1 Assistant Professor of Comparative Anatomy and Embryology, Department of Histology, Faculty of Paraveterinary Medicine, Ilam University, Ilam, Iran.

2 Associate Professor of Veterinary Anatomical Sciences, Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of ahvaz, Ahvaz, Iran.

3 Associate Professor of Medical Anatomical Sciences, Department of Basic Sciences, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran

4 Associate Professor of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz-Ahvaz-Iran.

5 Assistant Professor of Histology, Department of Basic Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of ahvaz, Ahvaz, Iran.



Background: MS is an autoimmune, inflammatory disease with demyelination and astrogliosis of the central nervous system that affects the brain and spinal cord. Its prevalence in women is three times that of men.
methods: This study was conducted on 42 male and female rats with an approximate weight of 300-350 grams in Shahid Chamran Faculty of Veterinary Medicine, Ahvaz. Material: The male sham group (received normal saline), the female sham group (received normal saline), the male MS group and the female MS group were divided. In order to inject ethidium bromide in the MS groups using a microinjector, rats were Ketamine (80 mg/kg) and xylazine (5 mg/kg) were anesthetized, 0.02% ethidium bromide and 20microliter normal saline were injected using a Hamilton syringe.
Results: The results of this study showed that the inflammation and degeneration of nerve cells increased in the MS groups, and this inflammation and degeneration was more in males, and the level of MBP expression in the MS group was reduced compared to other groups, which compared The two genders of this reduction in expression were almost the same, and the amount of GFAP expression in the MS group was increased compared to the other groups, which was higher in the female gender when comparing the two genders.
Conclusion: MS causes a decrease in the amount of myelin basic protein and an increase in the astrogliosis factor, which leads to the destruction of the myelin sheath and astrocytes and causes sensory and motor disabilities in people with MS.


Aref Nooraei [PumMed] [Google Scholar]

 Kaveh Khazaee [PumMed] [Google Scholar]

Marzieh Darvishi [PumMed] [Google Scholar]

 Zohreh Ghotbeddin [PumMed] [Google Scholar]

 Zahra Basir [PumMed] [Google Scholar]


Main Subjects

[1] Höftberger R, Lassmann H. Inflammatory demyelinating diseases of the central nervous system. Handb Clin Neurol. 2017;145:263-83. [DOI: 10.1016/B978-0-12-802395-2.00019-5] [PMID] [PMCID] [2] Fornasiero EF, Mandad S, Wildhagen H, Alevra M, Rammner B, Keihani S, et al. Precisely measured protein lifetimes in the mouse brain reveal differences across tissues and subcellular fractions. Nature communications. 2018;9(1):4230. [DOI: 10.1038/s41467-018-06519-0] [3] Martinsen V, Kursula P. Multiple sclerosis and myelin basic protein: Insights into protein disorder and disease. Amino Acids. 2022;54(1):99-109. [DOI: 10.1007/s00726-021-03111-7] [PMID] [PMCID] [4] Neumann B, Segel M, Chalut KJ, Franklin RJ. Remyelination and ageing: reversing the ravages of time. Mult Scler. 2019; 25(14):1835-41. [DOI: 10.1177/1352458519884006] [PMID] [PMCID] [5] Sofroniew MV. Astrogliosis. Cold Spring Harb Perspect Biol. 2014;7(2):a020420. [DOI: 10.1101/cshperspect.a020420] [PMID] [6] Mañé-Martínez MA, Olsson B, Bau L, Matas E, Cobo-Calvo Á, Andreasson U, et al. Glial and neuronal markers in cerebrospinal fluid in different types of multiple sclerosis. Mult Scler. 2015;21(5):550-61. [DOI: 10.1177/1352458514549397] [PMID] [PMCID] [7] Kassubek R, Gorges M, Schocke M, Hagenston VA, Huss A, Ludolph AC, et al. GFAP in early multiple sclerosis: A biomarker for inflammation. Neurosci Lett. 2017;657:166-70. [DOI: 10.1016/j.neulet.2017.07.050] [PMID] [8] De Armond SJ, Eng LF, Rubinstein LJ. The Application of filial Fibrillary Acidic (GFA) Protein Immunohistochemistry in Neurooncology: A Progress Report. Pathol Res Pract. 1980;168(4):374-94. [DOI: 10.1016/s0344-0338(80)80273-1] [PMID] [9] Raghavan R, Steart PV, Weller RO. Cell proliferation patterns in the diagnosis of astrocytomas, anaplastic astrocytomas and glioblastoma multiforme: a Ki‐67 study. Neuropathol Appl Neurobiol. 1990;16(2):123-33. [DOI: 10.1111/j.1365-2990. 1990.tb00941.x] [PMID] [10] Zhang N, Shang Z, Wang Z, Meng X, Li Z, Tian H, et al. Molecular pathological expression in malignant gliomas resected by fluorescein sodium-guiding under the YELLOW 560 nm surgical microscope filter. World J Surg Oncol. 2018;16(1):195. [DOI: 10.1186/s12957-018-1495-2] [PMID] [PMCID] [11] Bø L. The histopathology of grey matter demyelination in multiple sclerosis. Acta Neurol Scand Suppl. 2009;(189):51-7. [DOI: 10.1111/j.1600-0404.2009.01216.x] [PMID] [12] Ponath G, Park C, Pitt D. The role of astrocytes in multiple sclerosis. Front Immunol. 2018;9:217. [DOI: 10.3389/fimmu. 2018.00217] [PMID] [PMCID] [13] Brosnan CF, Raine CS. The astrocyte in multiple sclerosis revisited. Glia. 2013;61(4):453-65. [DOI: 10.1002/glia.22443] [PMID] [14] Levine JM, Reynolds R. Activation and proliferation of endogenous oligodendrocyte precursor cells during ethidium bromide-induced demyelination. Exp Neurol. 1999;160(2):333-47. [DOI: 10.1006/exnr.1999.7224] [PMID] [15] Barone FC, Hillegass LM, Price WJ, White RF, Lee EV, Feuerstein GZ, et al. Polymorphonuclear leukocyte infiltration into cerebral focal ischemic tissue: myeloperoxidase activity assay and histologic verification. J Neurosci Res. 1991;29(3):336-45. [DOI: 10.1002/jnr.490290309] [PMID] [16] Shirazi A, Golab F, Sanadgol N, Barati M, Mohammad Salehi R, Vahabzadeh G, et al. [Evaluation of the neurotrophic factors in animal model of myelin destruction induced by cuprizone in c57bl/6 mice (Persian)]. Shefaye Khatam. 2016;4(2):47-54. [DOI: 10.18869/acadpub.shefa.4.2.47] [17] Halliday GM, Cullen KM, Kril JJ, Harding AJ, Harasty J. Glial fibrillary acidic protein (GFAP) immunohistochemistry in human cortex: a quantitative study using different antisera. Neurosci Lett. 1996;209(1):29-32. [DOI: 10.1016/0304-3940(96)12592-1] [PMID] [18] Hamano K, Iwasaki N, Takeya T, Takita H. A quantitative analysis of rat central nervous system myelination using the immunohistochemical method for MBP. Brain Res Dev Brain Res. 1996;93(1-2):18-22. [DOI: 10.1016/0165-3806(96)00025-9] [PMID] [19] Yi W, Schlueter D, Wang X. Astrocytes in multiple sclerosis and experimental autoimmune encephalomyelitis: Star-shaped cells illuminating the darkness of CNS autoimmunity. Brain Behav Immun. 2019;80:10-24. [DOI: 10.1016/j.bbi.2019.05.029] [PMID] [20] Dunn SE, Gunde E, Lee H. Sex-based differences in multiple sclerosis (MS): part II: rising incidence of multiple sclerosis in women and the vulnerability of men to progression of this disease. Curr Top Behav Neurosci. 2015;26:57-86. [DOI: 10.1007/7854_2015_370] [PMID] [21] Kipp M, Beyer C. Impact of sex steroids on neuroinflammatory processes and experimental multiple sclerosis. Front Neuroendocrinol. 2009;30(2):188-200. [DOI: 10.1016/j.yfrne. 2009.04.004] [PMID] [22] Harp CT, Ireland S, Davis LS, Remington G, Cassidy B, Cravens PD, et al. Memory B cells from a subset of treatment‐naïve relapsing‐remitting multiple sclerosis patients elicit CD4+ T‐cell proliferation and IFN‐γ production in response to myelin basic protein and myelin oligodendrocyte glycoprotein. Eur J Immunol. 2010;40(10):2942-56. [DOI: 10.1002/eji.201040516] [PMID] [23] Jana M, Pahan K. Down-regulation of myelin gene expression in human oligodendrocytes by nitric oxide: implications for demyelination in multiple sclerosis. J Clin Cell Immunol. 2013;4:10.4172/2155-9899.1000157. [DOI: 10.4172/2155-9899.1000157] [PMID] [24] Boullerne, A. I., & Benjamins, J. A. (2006). Nitric oxide synthase expression and nitric oxide toxicity in oligodendrocytes. Antioxidants & redox signaling, 2006,8(5-6), 967-980. [DOI: 10.1089/ars.2006.8.96] [25] Calderon TM, Eugenin EA, Lopez L, Kumar SS, Hesselgesser J, Raine CS, et al. A role for CXCL12 (SDF-1α) in the pathogenesis of multiple sclerosis: regulation of CXCL12 expression in astrocytes by soluble myelin basic protein. J Neuroimmunol. 2006;177(1-2):27-39. [DOI: 10.1016/j.jneuroim.2006.05.003] [PMID] [26] Banik NL. Pathogenesis of myelin breakdown in demyelinating diseases: role of proteolytic enzymes. Crit Rev Neurobiol.
Nooraei A, et al. Expression Levels of MBP and GFAP Following the Induction of Neuronal Demyelination with Ethidium Bromide. JSMJ. 2024;
Journal of Medical Sciences
November & December 2024. Vol 22. No 5
1992;6(4):257-71. [PMID] [27] D’Souza CA, Moscarello MA. Differences in susceptibility of MBP charge isomers to digestion by stromelysin-1 (MMP-3) and release of an immunodominant epitope. Neurochemical research. 2006;31:1045-54. [DOI: 10.1007/s11064-006-9116-9] [28] Mastronardi FG, Moscarello MA. Molecules affecting myelin stability: a novel hypothesis regarding the pathogenesis of multiple sclerosis. J Neurosci Res. 2005;80(3):301-8. [DOI: 10.1002/jnr.20420] [PMID] [29] Sajad M, Zargan J, Chawla R, Umar S, Khan HA. Upregulation of CSPG3 accompanies neuronal progenitor proliferation and migration in EAE. J Mol Neurosci. 2011;43(3):531-40. [DOI: 10.1007/s12031-010-9476-0] [PMID] [30] Ludwin SK, Rao VT, Moore CS, Antel JP. Astrocytes in multiple sclerosis. Mult Scler. 2016;22(9):1114-24. [DOI: 10.1177/135 2458516643396] [PMID] [31] He F, Sun YE. Glial cells more than support cells? Int J Biochem Cell Biol. 2007;39(4):661-5. [DOI: 10.1016/j.biocel.2006. 10.022] [PMID] [32] Hatada I, Namihira M, Morita S, Kimura M, Horii T, Nakashima K. Astrocyte-specific genes are generally demethylated in neural precursor cells prior to astrocytic differentiation. PLoS One. 2008;3(9):e3189. [DOI: 10.1371/journal.pone.0003189] [PMID] [33] Sun D, Jakobs TC. Structural remodeling of astrocytes in the injured CNS. Neuroscientist. 2012;18(6):567-88. [DOI: 10.1177/1073858411423441] [PMID] [PMCID] [34] [34] Abdelhak A, Hottenrott T, Morenas-Rodríguez E, Suárez-Calvet M, Zettl UK, Haass C, et al. Glial activation markers in CSF and serum from patients with primary progressive multiple sclerosis: potential of serum GFAP as disease severity marker? Front Neurol. 2019;10:280. [DOI: 10.3389/fneur. 2019.00280] [PMID] [PMCID] [35] Abdelhak, A., Huss, A., Kassubek, J., Tumani, H., & Otto, M. Serum GFAP as a biomarker for disease severity in multiple sclerosis. Scientific reports,2018, 8(1), 14798. [DOI:10.1038/ s41598-018-33158-8]. [36] Norkute A, Hieble A, Braun A, Johann S, Clarner T, Baumgartner W, et al. Cuprizone treatment induces demyelination and astrocytosis in the mouse hippocampus. J Neurosci Res. 2009;87(6):1343-55. [DOI: 10.1002/jnr.21946] [PMID] [37] Sriram K, Benkovic SA, Hebert MA, Miller DB, O'Callaghan JP. Induction of gp130-related cytokines and activation of JAK2/STAT3 pathway in astrocytes precedes up-regulation of glial fibrillary acidic protein in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine model of neurodegeneration: key signaling pathway for astrogliosis in vivo? J Biol Chem. 2004;279(19):19936-47. [DOI: 10.1074/jbc.M309304200] [PMID] [38] Rozovsky I, Wei M, Stone DJ, Zanjani H, Anderson CP, Morgan TE, et al. Estradiol (E2) enhances neurite outgrowth by repressing glial fibrillary acidic protein expression and reorganizing laminin. Endocrinology. 2002;143(2):636-46. [DOI: 10.1210/endo.143.2.8615] [PMID]