Evaluation of apamin effects on myelination process in C57BL/6 mice model of multiple sclerosis

Maedeh Mohammadi-Rad , Nazem Ghasemi, Mehdi Aliomrani


Multiple sclerosis (MS) is a demyelinating disease that causes chronic inflammation in the central nervous system. The aim of this study was to investigate the effects of apamin administration on myelination process. MS was induced by feeding cuprizone pellets (0.2%) for 6 weeks (demyelination phase) followed by normal feeding for additional 2 weeks (remyelination phase). Briefly, C57BL/6 male mice were randomly divided into six groups. Group 1, received the regular food pellets. Group 2 contained two subgroups of 6 animals each (n = 2 × 6). First group received cuprizone for 6 weeks and the sacrificed while the second group after 6 weeks of cuprizone, received no treatment for additional 2 weeks. Group 3 (co-treatment group) was composed of two subgroups of 6 animals each (n = 2 × 6). Both subgroups received apamin (100 μg/kg) intraperitoneally twice a week for 6 weeks. First subgroup terminated at this time and the second subgroup was fed normal diet for two additional weeks. Group 4 (post-treatment, n = 6) received apamin (100 μg/kg) intraperitoneally twice a week for 2 weeks after cuprizone secession. Groups 5 and 6 (vehicle, n = 6 in each group) received phosphate buffered saline as the vehicle of apamin during demyelination and remyelination phase. At the end of each phase, mice were deeply anesthetized and perfused. Groups 5 and 6 (vehicle) received PBS as the vehicle during both phases. Mice were anesthetized, perfused with PBS through their heart, and their brains were removed. Brain sections stained with luxol fast blue and the images were analyzed. Apamin co-treatment significantly increased the myelin content as compared to the cuprizone group. Also, mild elevation in the myelinated areas was observed with apamin post-treatment in comparison with remyelination phase. Our results revealed that apamin prevents myelin destruction more significantly as compared to remyelination process. This observation explains the possible role of apamin in inhibiting   the activation of the microglia cells than stimulation of the oligodendrocytic precursor cells.


Apamin; C57BL/6; Cuprizone; Multiple sclerosis; Myelin.

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Olsson T, Barcellos LF, Alfredsson L. Interactions between genetic, lifestyle and environmental risk factors for multiple sclerosis. Nat Rev Neurol. 2017;13(1):25-36.

Aliomrani M, Sahraian MA, Shirkhanloo H, Sharifzadeh M, Khoshayand MR, Ghahremani MH. Correlation between heavy metal exposure and GSTM1 polymorphism in Iranian multiple sclerosis patients. Neurol Sci. 2017;38(7):1271-1278.

Rezapour-Firouzi S, Shahabi S, Mohammadzadeh A, Tehrani AA, Kheradmand F, Mazloomi E.The potential effects of hemp seed/evening primrose oils on the mammalian target of rapamycin complex 1 and interferon-gamma genes expression in experimental autoimmune encephalomyelitis. Res Pharm Sci. 2018;13(6):523-532.

Gold R, Wolinsky JS. Pathophysiology of multiple sclerosis and the place of teriflunomide. Acta Neurol Scand. 2011;124(2):75-84.

Aliomrani M, Sahraian MA, Shirkhanloo H, Sharifzadeh M, Khoshayand MR, Ghahremani MH. Blood concentrations of cadmium and lead in multiple sclerosis patients from Iran. Iran J Pharm Res. 2016;15(4):825-833.

Greenstein JI. Current concepts of the cellular and molecular pathophysiology of multiple sclerosis. Dev Neurobiol. 2007;67(9):1248-1265.

Dawson MR, Levine JM, Reynolds R. NG2‐expressing cells in the central nervous system: are they oligodendroglial progenitors? J Neurosci Res. 2000;61(5):471-479.

Eze OBL, Nwodo OFC, Ogugua VN. Therapeutic effect of honey bee venom. Res J Pharm Biol Chem Sci. 2016;4(1):48-53.

Khoei NS, Atashpaz S, Ghabili K, Khoei NS,Omidi Y. Melittin and hyaluronidase compound derived from Bee venom for the treatment of multiple sclerosis. J Med Hypotheses Ideas. 2009;3(3):24-28.

Moreno M, Giralt E. Three valuable peptides from bee and wasp venoms for therapeutic and biotechnological use: melittin, apamin and mastoparan. Toxins (Basel). 2015;7(4):1126-1150.

Voos P, Yazar M, Lautenschläger R, Rauh O, Moroni A, Thiel G. The small neurotoxin apamin blocks not only small conductance Ca2+ activated K+ channels (SK type) but also the voltage dependent Kv1. 3 channel. Eur Biophys J. 2017;46(6):517-523.

Shimpi R, Chaudhari P, Deshmukh R, Devare S, Bagad Y, Bhurat M. A review: pharmacotherapeutics of bee venom. Word J Pharm Pharm Sci. 2016;5(7):656-567.

Gage GJ, Kipke DR, Shain W. Whole animal perfusion fixation for rodents. J Vis Exp. 2012(65). pii: 3564.

McCormick JB. Apparatus and method for preparing tissue samples for histological examination.United State Patents US7771992B2; 2010. http://www.freepatentsonline.com/7771992.html.

Laule C, Leung E, Li DK, Traboulsee AL, Paty DW, MacKay AL, et al. Myelin water imaging in multiple sclerosis: quantitative correlations with histopathology. Mult Scler. 2006;12(6):747-753.

Ruifrok AC, Johnston DA. Quantification of histochemical staining by color deconvolution. Anal Quant Cytol Histol. 2001;23(4):291-299.

Zhen W, Liu A, Lu J, Zhang W, Tattersall D, Wang J. An alternative cuprizone-induced demyelination and remyelination mouse model. ASN Neuro. 2017;9(4):1-9.

Morell P, Barrett CV, Mason JL, Toews AD, Hostettler JD, Knapp GW, et al. Gene expression in brain during cuprizone-induced demyelination and remyelination. Mol Cell Neurosci. 1998; 12(4-5):220-227.

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

Hochstrasser T, Exner GL, Nyamoya S, Schmitz C, Kipp M. Cuprizone-containing pellets are less potent to induce consistent demyelination in the corpus callosum of C57BL/6 mice. J Mol Neurosci. 2017;61(4):617-624.

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

Gudi V, Gingele S, Skripuletz T, Stangel M. Glial response during cuprizone-induced de-and remyelination in the CNS: lessons learned.Front Cell Neurosci. 2014;8:73-96.

Alvarez-Fischer D, Noelker C, Vulinović F, Grünewald A, Chevarin C, Klein C, et al.Bee venom and its component apamin as neuroprotective agents in a Parkinson disease mouse model. PLoS One. 2013;8(4):e61700.

Dempsey CE, Sessions RB, Lamble NV,Campbell SJ. The asparagine-stabilized β-turn of apamin: contribution to structural stability from dynamics simulation and amide hydrogen exchange analysis. Biochemistry. 2000;39(51):15944-15952.

Silva J, Monge-Fuentes V, Gomes F, Lopes K, dos Anjos L, Campos G, et al. Pharmacological alternatives for the treatment of neurodegenerative disorders: wasp and bee venoms and their components as new neuroactive tools. Toxins (Basel). 2015;7(8):3179-3209.

Kim SJ, Park JH, Kim K-H, Lee W-R, Pak SC, Han S-M, et al. The protective effect of apamin on LPS/fat-induced atherosclerotic mice. Evid Based Complement Alternat Med. 2012;2012. Article ID 305454, 10 pages.

Ferreira R, Schlichter LC. Selective activation of KCa3. 1 and CRAC channels by P2Y2 receptors promotes Ca(2+) signaling, store refilling and migration of rat microglial cells. PLoS One. 2013;8(4):e62345.

Fordyce CB, Jagasia R, Zhu X, Schlichter LC. Microglia Kv1. 3 channels contribute to their ability to kill neurons. J Neurosci. 2005;25(31):7139-7149.

Benton DC, Garbarg M, Moss GW. The relationship between functional inhibition and binding for K(Ca)2 channel blockers. PloS One. 2013;8(9):e73328.

Liu H, Liu J, Xu E, Tu G, Guo M, Liang S, et al. Human immunodeficiency virus protein Tat induces oligodendrocyte injury by enhancing outward K+ current conducted by KV1.3. Neurobiol Dis. 2017;97(Pt A):1-10.


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