Attenuation of Magnesium Sulfate on CoCl_2-Induced Cell Death by Activating ERK1/2/MAPK and Inhibiting HIF-1α via Mitochondrial Apoptotic Signaling Suppression in a Neuronal Cell Line




Chih-Yang Huang;You-Liang Hsieh;Da-Tong Ju;Chien-Chung Lin;Chia-Hua Kuo;Yi-Fan Liou;Tsung-Jung Ho;Chang-Hai Tsai;Fuu-Jen Tsai;Jing-Ying Lin

Key Words

apoptosis ; Bcl-2 family ; CoCl_2, ERK1/2 ; MgSO_4 ; mitochondria


The Chinese Journal of Physiology

Volume or Term/Year and Month of Publication

58卷4期(2015 / 08 / 01)

Page #

244 - 253

Content Language


English Abstract

Magnesium sulfate (MgSO_4) ameliorates hypoxia/ischemia-induced neuronal apoptosis in a rat model. This study aimed to investigate the mechanisms governing the anti-apoptotic effect of MgSO_4 on cobalt chloride (CoCl_2)-exposed NB41A3 mouse neuroblastoma cells. MgSO_4 increased the viability of NB41A3 cells treated with CoCl_2 in a dose-dependent manner. MgSO_4 treatment was shown to lead to an increase in the anti-apoptotic Bcl-2 family proteins, with a concomitant decrease in the pro-apoptotic proteins. MgSO_4 also attenuated the CoCl2-induced disruption of mitochondrial membrane potential (ΔΨm) and reduced the release of cytochrome c form the mitochondria to the cytosol. Furthermore, exposure to CoCl2 caused activation of the hypoxia-inducible factor 1α (HIF-1α). On the other hand, MgSO_4 markedly reduced CoCl_2-induced HIF-1α activation and suppressed HIF-1α downstream protein BNIP3. MgSO_4 treatment induced ERK1/2 activation and attenuated CoCl_2-induced activation of p38 and JNK. Addition of the ERK1/2 inhibitor U0126 significantly reduced the ability of MgSO_4 to protect neurons from CoCl_2-induced mitochondrial apoptotic events. However, incubation of cultures with the p38 and JNK inhibitors did not significantly affect MgSO_4-mediated neuroprotection. MgSO_4 appears to suppress CoCl_2-induced NB41A3 cell death by activating ERK1/2/ MAPK pathways, which further modulates the role of Bcl-2 family proteins and mitochondria in NB41A3 cells. Our data suggest that MgSO_4 may act as a survival factor that preserves mitochondrial integrity and inhibits apoptotic pathways.

Topic Category 醫藥衛生 > 基礎醫學
  1. Huang, C.Y.,Liou, Y.F.,Chung, S.Y.,Lin, W.Y.,Jong, G.P.,Kuo, C.H.,Tsai, F.J.,Cheng, Y.C.,Cheng, F.C.,Lin, J.Y.(2010).Role of ERK signaling in the neuroprotective efficacy of magnesium sulfate treatment during focal cerebral ischemia in the gerbil cortex.Chinese J. Physiol.,53,299-309.
  2. Xiang, Q.L.,Chen, Y.,Lin, G.P.,Tan, Z.,Wang, T.H.(2006).Effects of selective estrogen receptor modulator raloxifene plus 17-beta estradiol in aorta and mammary gland of female experimental atherosclerosis rabbits and possible involvement of ERK signal transduction pathway.Chinese J. Physiol.,49,132-140.
  3. Aminova, L.R.,Chavez, J.C.,Lee, J.,Ryu, H.,Kung, A.,LaManna, J.C.,Ratan, R.R.(2005).Prosurvival and prodeath effects of hypoxiainducible factor-1α stabilization in a murine hippocampal cell line.J. Biol. Chem.,280,3996-4003.
  4. Barone, F.C.,Irving, E.A.,Ray, A.M.,Lee, J.C.,Kassis, S.,Kumar, S.,Badger, A.M.,Legos, J.J.,Erhardt, J.A.,Ohlstein, E.H.,Hunter, A.J.,Harrison, D.C.,Philpott, K.,Smith, B.R.,Adams, J.L.,Parsons, A.A.(2001).Inhibition of p38 mitogen-activated protein kinase provides neuroprotection in cerebral focal ischemia.Med. Res. Rev.,21,129-145.
  5. Bergeron, M.,Yu, A.Y.,Solway, K.E.,Semenza, G.L.,Sharp, F.R.(1999).Induction of hypoxia-inducible factor-1 (HIF-1) and its target genes following focal ischaemia in rat brain.Eur. J. Neurosci.,11,4159-4170.
  6. Chachami, G.,Simos, G.,Hatziefthimiou, A.,Bonanou, S.,Molyvdas, P.A.,Paraskeva, E.(2004).Cobalt induces hypoxia-inducible factor-1α expression in airway smooth muscle cells by a reactive oxygen species- and PI3K-dependent mechanism.Am. J. Respir. Cell Mol. Biol.,31,544-551.
  7. Cheng, C.Y.,Lin, J.G.,Su, S.Y.,Tang, N.Y.,Kao, S.T.,Hsieh, C.L.(2014).Electroacupuncture-like stimulation at Baihui and Dazhui acupoints exerts neuroprotective effects through activation of the brain-derived neurotrophic factor-mediated MEK1/2/ERK1/2/p90RSK/bad signaling pathway in mild transient focal cerebral ischemia in rats.BMC Complement. Altern. Med.,14,92.
  8. Doyle, L.W.(2012).Antenatal magnesium sulfate and neuroprotection..Curr. Opin. Pediatr,24,154-159.
  9. Ellerby, L.M.,Ellerby, H.M.,Park, S.M.,Holleran, A.L.,Murphy, A.N.,Fiskum, G.,Kane, D.J.,Testa, M.P.,Kayalar, C.,Bredesen, D.E.(1996).Shift of the cellular oxidation-reduction potential in neural cells expressing Bcl-2.J. Neurochem.,67,1259-1267.
  10. Fan, M.,Goodwin, M.,Vu, T.,Brantley-Finley, C.,Gaarde, W.A.,Chambers, T.C.(2000).Vinblastine-induced phosphorylation of Bcl-2 and Bcl-XL is mediated by JNK and occurs in parallel with inactivation of the Raf-1/MEK/ERK cascade.J. Biol. Chem.,275,29980-29985.
  11. Ferrer, I.,Ballabriga, J.,Pozas, E.(1997).Transient forebrain ischemia in the adult gerbil is associated with a complex c-Jun response.Neuroreport,8,2483-2487.
  12. Heidenreich, K.A.,Kummer, J.L.(1996).Inhibition of p38 mitogenactivated protein kinase by insulin in cultured fetal neurons.J. Biol. Chem.,271,9891-9894.
  13. Helton, R.,Cui, J.,Scheel, J.R.,Ellison, J.A.,Ames, C.,Gibson, C.,Blouw, B.,Ouyang, L.,Dragatsis, I.,Zeitlin, S.,Johnson, R.S.,Lipton, S.A.,Barlow, C.(2005).Brain-specific knock-out of hypoxiainducible factor-1α reduces rather than increases hypoxic-ischemic damage.J. Neurosci.,25,4099-4107.
  14. Huang, C.Y.,Liou, Y.F.,Chung, S.Y.,Pai, P.Y.,Kan, C.B.,Kuo, C.H.,Tsai, C.H.,Tsai, F.J.,Chen, J.L.,Lin, J.Y.(2010).Increased expression of glucose transporter 3 in gerbil brains following magnesium sulfate treatment and focal cerebral ischemic injury.Cell Biochem. Funct.,28,313-320.
  15. Irving, E.A.,Bamford, M.(2002).Role of mitogen- and stress-activated kinases in ischemic injury.J. Cereb. Blood Flow Metab.,66,631-647.
  16. Jin, K.L.,Mao, X.O.,Nagayama, T.,Goldsmith, P.C.,Greenberg, D.A.(2000).Induction of vascular endothelial growth factor and hypoxia-inducible factor-1α by global ischemia in rat brain.Neuroscience,99,577-585.
  17. Jones, N.M.,Bergeron, M.(2001).Hypoxic preconditioning induces changes in HIF-1 target genes in neonatal rat brain.J. Cereb. Blood Flow Metab.,21,1105-1114.
  18. Kitagawa, K.(2007).CREB and cAMP response element-mediated gene expression in the ischemic brain.FEBS J.,274,3210-3217.
  19. Kitagawa, K.,Matsumoto, M.,Tsujimoto, Y.,Ohtsuki, T.,Kuwabara, K.,Matsushita, K.,Yang, G.,Tanabe, H.,Martinou, J.C.,Hori, M.,Yanagihara, T.(1998).Amelioration of hippocampal neuronal damage after global ischemia by neuronal overexpression of BCL-2 in transgenic mice.Stroke,29,2616-2621.
  20. Lee, J.H.,Choi, S.H.,Baek, M.W.,Kim, M.H.,Kim, H.J.,Kim, S.H.,Oh, S.J.,Park, H.J.,Kim, W.J.,Jung, J.Y.(2013).CoCl2 induces apoptosis through the mitochondria- and death receptor-mediated pathway in the mouse embryonic stem cells.Mol. Cell Biochem.,379,133-140.
  21. Lin, J.Y.,Chung, S.Y.,Lin, M.C.,Cheng, F.C.(2002).Effects of magnesium sulfate on energy metabolites and glutamate in the cortex during focal cerebral ischemia and reperfusion in the gerbil monitored by a dual-probe microdialysis technique.Life Sci.,71,803-811.
  22. Liu, Y.,Chen, L.(2012).Mechanisms in adaptation to brain ischemia by ischemic preconditioning.Adapt. Med.,4,20-26.
  23. Mayer, M.L.,Westbrook, G.L.,Guthrie, P.B.(1984).Voltage-dependent block by Mg2+ of NMDA responses in spinal cord neurones.Nature,309,261-263.
  24. Merker, H.J.,Gunther, T.,Hollriegl, V.,Vormann, J.,Schumann, K.(1996).Lipid peroxidation and morphology of rat testis in magnesium deficiency.Andrologia,28,43-51.
  25. Muir, K.W.(2002).Magnesium in stroke treatment.Postgrad. Med. J.,78,641-645.
  26. Nakka, V.P.,Gusain, A.,Mehta, S.L.,Raghubir, R.(2008).Molecular mechanisms of apoptosis in cerebral ischemia: multiple neuroprotective opportunities.Mol. Neurobiol.,37,7-38.
  27. Ozawa, H.,Shioda, S.,Dohi, K.,Matsumoto, H.,Mizushima, H.,Zhou, C.J.,Funahashi, H.,Nakai, Y.,Nakajo, S.,Matsumoto, K.(1999).Delayed neuronal cell death in the rat hippocampus is mediated by the mitogen-activated protein kinase signal transduction pathway.Neurosci. Lett.,262,57-60.
  28. Piret, J.P.,Mottet, D.,Raes, M.,Michiels, C.(2002).Is HIF-1α a proor an anti-apoptotic protein?.Biochem. Pharmacol.,64,889-892.
  29. Plotnikov, A.,Zehorai, E.,Procaccia, S.,Seger, R.(2011).The MAPK cascades: signaling components, nuclear roles and mechanisms of nuclear translocation.Biochim. Biophys. Acta,1813,1619-1633.
  30. Ratcliffe, P.J.,O'Rourke, J.F.,Maxwell, P.H.,Pugh, C.W.(1998).Oxygen sensing, hypoxia-inducible factor-1 and the regulation of mammalian gene expression.J. Exp. Biol.,201,1153-1162.
  31. Rayssiguier, Y.,Gueux, E.,Bussiere, L.,Durlach, J.,Mazur, A.(1993).Dietary magnesium affects susceptibility of lipoproteins and tissues to peroxidation in rats.J. Am. Coll. Nutr,12,133-137.
  32. Regan, R.F.,Jasper, E.,Guo, Y.,Panter, S.S.(1998).The effect of magnesium on oxidative neuronal injury in vitro.J. Neurochem.,70,77-85.
  33. Sameshima, H.,Ikenoue, T.(2002).Effect of long-term, postasphyxial administration of magnesium sulfate on immunostaining of microtubule-associated protein-2 and activated caspase-3 in 7-day-old rat brain.J. Soc. Gynecol. Investig.,9,203-209.
  34. Semenza, G.L.(2001).Hypoxia-inducible factor 1: oxygen homeostasis and disease pathophysiology.Trends Mol. Med.,7,345-350.
  35. Semenza, G.L.(2007).Life with oxygen.Science,318,62-64.
  36. Sharp, F.R.,Ran, R.,Lu, A.,Tang, Y.,Strauss, K.I.,Glass, T.,Ardizzone, T.,Bernaudin, M.(2004).Hypoxic preconditioning protects against ischemic brain injury.NeuroRx,1,26-35.
  37. Solaroglu, I.,Kaptanoglu, E.,Okutan, O.,Beskonakli, E.,Attar, A.,Kilinc, K.(2005).Magnesium sulfate treatment decreases caspase-3 activity after experimental spinal cord injury in rats.Surg. Neurol.,64(Suppl 2),S17-S21.
  38. Strohm, C.,Barancik, T.,Bruhl, M.L.,Kilian, S.A.,Schaper, W.(2000).Inhibition of the ER-kinase cascade by PD98059 and UO126 counteracts ischemic preconditioning in pig myocardium.J. Cardiovasc. Pharmacol.,36,218-229.
  39. Torii, S.,Kobayashi, K.,Takahashi, M.,Katahira, K.,Goryo, K.,Matsushita, N.,Yasumoto, K.,Fujii-Kuriyama, Y.,Sogawa, K.(2009).Magnesium deficiency causes loss of response to intermittent hypoxia in paraganglion cells.J. Biol. Chem.,284,19077-19089.
  40. Turkyilmaz, C.,Turkyilmaz, Z.,Atalay, Y.,Soylemezoglu, F.,Celasun, B.(2002).Magnesium pre-treatment reduces neuronal apoptosis in newborn rats in hypoxia-ischemia.Brain Res.,955,133-137.
  41. Wiener, C.M.,Booth, G.,Semenza, G.L.(1996).In vivo expression of mRNAs encoding hypoxia-inducible factor 1.Biochem. Biophys. Res. Commun.,225,485-488.
  42. Yin, X.M.,Luo, Y.,Cao, G.,Bai, L.,Pei, W.,Kuharsky, D.K.,Chen, J.(2002).Bid-mediated mitochondrial pathway is critical to ischemic neuronal apoptosis and focal cerebral ischemia.J. Biol. Chem.,277,42074-42081.
  43. Zhang, F.,Chen, J.(2008).Leptin protects hippocampal CA1 neurons against ischemic injury.J. Neurochem.,107,578-587.
  44. Zheng, S.,Zuo, Z.(2004).Isoflurane preconditioning induces neuroprotection against ischemia via activation of P38 mitogenactivated protein kinases.Mol. Pharmacol.,65,1172-1180.
  45. Zou, W.,Yan, M.,Xu, W.,Huo, H.,Sun, L.,Zheng, Z.,Liu, X.(2001).Cobalt chloride induces PC12 cells apoptosis through reactive oxygen species and accompanied by AP-1 activation.J. Neurosci. Res.,64,646-653.
Times Cited
  1. 簡鸝慧、詹貴惠、楊明達、林書丞(2018)。單次甜菜鹼增補對力竭性耐力運動後細胞凋亡、氧化壓力及有氧能力的影響。大專體育學刊,20(4),373-383。