Molecular Neurobiology

, Volume 53, Issue 1, pp 695–705 | Cite as

Atorvastatin Protects NSC-34 Motor Neurons Against Oxidative Stress by Activating PI3K, ERK and Free Radical Scavenging

  • Seok-Ho Lee
  • Na-Young Choi
  • Hyun-Jeung Yu
  • Jinse Park
  • Hojin Choi
  • Kyu-Yong Lee
  • Yong-Min Huh
  • Young Joo Lee
  • Seong-Ho Koh
Article
First Online:
Received:
Accepted:

Abstract

Although statins, or hydroxymethylglutaryl coenzyme A (HMG-Co A) reductase inhibitors, are generally used to decrease levels of circulating cholesterol, they have also been reported to have neuroprotective effects through various mechanisms. However, recent results have indicated that they may be harmful in patients with amyotrophic lateral sclerosis (ALS). In this study, we investigate whether atorvastatin protects motor neuron-like cells (NSC-34D) from oxidative stress. To evaluate the effects of atorvastatin or hydrogen peroxide or both on NSC-34D cells, the cells were treated with various combinations of these agents. To evaluate the viability of the cells, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays and trypan blue staining were performed. Levels of free radicals and intracellular signaling proteins were evaluated using the fluorescent probe 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) and Western blotting, respectively. Atorvastatin protected NSC-34D cells against oxidative stress in a concentration-dependent manner. This neuroprotective effect of atorvastatin was blocked by LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor and by FR180204, a selective extracellular signal-related kinase (ERK) inhibitor. Atorvastatin treatment increased the expression levels of p85αPI3K, phosphorylated Akt, phosphorylated glycogen synthase kinase-3β, phosphorylated ERK, and Bcl-2, which are proteins related to survival. Furthermore, atorvastatin decreased the levels of cytosolic cytochrome C, Bax, cleaved caspase-9, and cleaved caspase-3, which are associated with death in oxidative stress-injured NSC-34D cells. We conclude that atorvastatin has a protective effect against oxidative stress in motor neurons by activating the PI3K and ERK pathways as well as by scavenging free radicals. These findings indicate that statins could help protect motor neurons from oxidative stress.

Keywords

Statin Amyotrophic lateral sclerosis Phosphatidylinositol 3-kinase Extracellular signal-related kinase 
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Notes

Acknowledgments

This work was supported by a grant from the Korea Research Foundation (2012R1A1B3000473) and a grant from the NanoBio R&D Program of the Korea Science and Engineering Foundation, funded by the Ministry of Education, Science and Technology (2007–04717).

Conflict of Interest

None.

References

  1. 1.
    Stein EA (2002) Management of dyslipidemia in the high-risk patient. Am Heart J 144(6 Suppl):S43–S50. doi: 10.1067/mhj.2002.130302 CrossRefPubMedGoogle Scholar
  2. 2.
    Sakabe K, Fukuda N, Wakayama K, Nada T, Shinohara H, Tamura Y (2004) Lipid-altering changes and pleiotropic effects of atorvastatin in patients with hypercholesterolemia. Am J Cardiol 94(4):497–500. doi: 10.1016/j.amjcard.2004.04.067 CrossRefPubMedGoogle Scholar
  3. 3.
    Wahner AD, Bronstein JM, Bordelon YM, Ritz B (2008) Statin use and the risk of Parkinson disease. Neurology 70(16 Pt 2):1418–1422. doi: 10.1212/01.wnl.0000286942.14552.51 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Eckert GP, Wood WG, Muller WE (2005) Statins: drugs for Alzheimer’s disease? J Neural Transm 112(8):1057–1071. doi: 10.1007/s00702-004-0273-1 CrossRefPubMedGoogle Scholar
  5. 5.
    Neuhaus O, Hartung HP (2007) Evaluation of atorvastatin and simvastatin for treatment of multiple sclerosis. Expert Rev Neurother 7(5):547–556. doi: 10.1586/14737175.7.5.547 CrossRefPubMedGoogle Scholar
  6. 6.
    Wible EF, Laskowitz DT (2010) Statins in traumatic brain injury. Neurother : J Am Soc Exp Neurother 7(1):62–73. doi: 10.1016/j.nurt.2009.11.003 CrossRefGoogle Scholar
  7. 7.
    Edwards IR, Star K, Kiuru A (2007) Statins, neuromuscular degenerative disease and an amyotrophic lateral sclerosis-like syndrome: an analysis of individual case safety reports from vigibase. Drug Saf : Int J Med Toxicol Drug Exp 30(6):515–525CrossRefGoogle Scholar
  8. 8.
    Colman E, Szarfman A, Wyeth J, Mosholder A, Jillapalli D, Levine J, Avigan M (2008) An evaluation of a data mining signal for amyotrophic lateral sclerosis and statins detected in FDA’s spontaneous adverse event reporting system. Pharmacoepidemiol Drug Saf 17(11):1068–1076. doi: 10.1002/pds.1643 CrossRefPubMedGoogle Scholar
  9. 9.
    Zinman L, Sadeghi R, Gawel M, Patton D, Kiss A (2008) Are statin medications safe in patients with ALS? Amyotroph Lateral Scler : Off Publ World Fed Neurol Res Group Mot Neuron Dis 9(4):223–228. doi: 10.1080/17482960802031092 CrossRefGoogle Scholar
  10. 10.
    Drory VE, Bronipolsky T, Artamonov I, Nefussy B (2008) Influence of statins treatment on survival in patients with amyotrophic lateral sclerosis. J Neurol Sci 273(1–2):81–83. doi: 10.1016/j.jns.2008.06.022 CrossRefPubMedGoogle Scholar
  11. 11.
    Iwamoto K, Yoshii Y, Ikeda K (2009) Atorvastatin treatment attenuates motor neuron degeneration in Wobbler mice. Amyotroph Lateral Scler: Off Publ World Fed Neurol Res Group Mot Neuron Dis 10(5–6):405–409. doi: 10.3109/17482960902870993 CrossRefGoogle Scholar
  12. 12.
    Cashman NR, Durham HD, Blusztajn JK, Oda K, Tabira T, Shaw IT, Dahrouge S, Antel JP (1992) Neuroblastoma x spinal cord (NSC) hybrid cell lines resemble developing motor neurons. Dev Dyn : Off Publ Am Assoc Anat 194(3):209–221. doi: 10.1002/aja.1001940306 CrossRefGoogle Scholar
  13. 13.
    Eggett CJ, Crosier S, Manning P, Cookson MR, Menzies FM, McNeil CJ, Shaw PJ (2000) Development and characterisation of a glutamate-sensitive motor neuron cell line. J Neurochem 74(5):1895–1902CrossRefPubMedGoogle Scholar
  14. 14.
    Hemendinger RA, Armstrong EJ 3rd, Radio N, Brooks BR (2012) Neurotoxic injury pathways in differentiated mouse motor neuron-neuroblastoma hybrid (NSC-34D) cells in vitro—limited effect of riluzole on thapsigargin, but not staurosporine, hydrogen peroxide and homocysteine neurotoxicity. Toxicol Appl Pharmacol 258(2):208–215. doi: 10.1016/j.taap.2011.10.022 CrossRefPubMedGoogle Scholar
  15. 15.
    Cantrell DA (2001) Phosphoinositide 3-kinase signalling pathways. J Cell Sci 114(Pt 8):1439–1445PubMedGoogle Scholar
  16. 16.
    Cantley LC (2002) The phosphoinositide 3-kinase pathway. Science 296(5573):1655–1657. doi: 10.1126/science.296.5573.1655 CrossRefPubMedGoogle Scholar
  17. 17.
    Cookson MR, Shaw PJ (1999) Oxidative stress and motor neurone disease. Brain Pathol 9(1):165–186CrossRefPubMedGoogle Scholar
  18. 18.
    Kim YS, Noh MY, Kim JY, Yu HJ, Kim KS, Kim SH, Koh SH (2013) Direct GSK-3beta inhibition enhances mesenchymal stromal cell migration by increasing expression of beta-PIX and CXCR4. Mol Neurobiol 47(2):811–820. doi: 10.1007/s12035-012-8393-3 CrossRefPubMedGoogle Scholar
  19. 19.
    Vanhaesebroeck B, Waterfield MD (1999) Signaling by distinct classes of phosphoinositide 3-kinases. Exp Cell Res 253(1):239–254. doi: 10.1006/excr.1999.4701 CrossRefPubMedGoogle Scholar
  20. 20.
    Smith MA, Perry G, Richey PL, Sayre LM, Anderson VE, Beal MF, Kowall N (1996) Oxidative damage in Alzheimer’s. Nature 382(6587):120–121. doi: 10.1038/382120b0 CrossRefPubMedGoogle Scholar
  21. 21.
    van der Most PJ, Dolga AM, Nijholt IM, Luiten PG, Eisel UL (2009) Statins: mechanisms of neuroprotection. Prog Neurobiol 88(1):64–75. doi: 10.1016/j.pneurobio.2009.02.002 CrossRefPubMedGoogle Scholar
  22. 22.
    Bosel J, Gandor F, Harms C, Synowitz M, Harms U, Djoufack PC, Megow D, Dirnagl U, Hortnagl H, Fink KB, Endres M (2005) Neuroprotective effects of atorvastatin against glutamate-induced excitotoxicity in primary cortical neurones. J Neurochem 92(6):1386–1398. doi: 10.1111/j.1471-4159.2004.02980.x CrossRefPubMedGoogle Scholar
  23. 23.
    Wang J, Tokoro T, Matsui K, Higa S, Kitajima I (2005) Pitavastatin at low dose activates endothelial nitric oxide synthase through PI3K-AKT pathway in endothelial cells. Life Sci 76(19):2257–2268. doi: 10.1016/j.lfs.2004.12.003 CrossRefPubMedGoogle Scholar
  24. 24.
    Dimmeler S, Aicher A, Vasa M, Mildner-Rihm C, Adler K, Tiemann M, Rutten H, Fichtlscherer S, Martin H, Zeiher AM (2001) HMG-CoA reductase inhibitors (statins) increase endothelial progenitor cells via the PI 3-kinase/Akt pathway. J Clin Invest 108(3):391–397. doi: 10.1172/JCI13152 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Zipp F, Waiczies S, Aktas O, Neuhaus O, Hemmer B, Schraven B, Nitsch R, Hartung HP (2007) Impact of HMG-CoA reductase inhibition on brain pathology. Trends Pharmacol Sci 28(7):342–349. doi: 10.1016/j.tips.2007.05.001 CrossRefPubMedGoogle Scholar
  26. 26.
    Wagner AH, Kohler T, Ruckschloss U, Just I, Hecker M (2000) Improvement of nitric oxide-dependent vasodilatation by HMG-CoA reductase inhibitors through attenuation of endothelial superoxide anion formation. Arterioscler Thromb Vasc Biol 20(1):61–69CrossRefPubMedGoogle Scholar
  27. 27.
    Weitz-Schmidt G, Welzenbach K, Brinkmann V, Kamata T, Kallen J, Bruns C, Cottens S, Takada Y, Hommel U (2001) Statins selectively inhibit leukocyte function antigen-1 by binding to a novel regulatory integrin site. Nat Med 7(6):687–692. doi: 10.1038/89058 CrossRefPubMedGoogle Scholar
  28. 28.
    Wood WG, Igbavboa U, Muller WE, Eckert GP (2013) Statins, Bcl-2, and apoptosis: cell death or cell protection? Mol Neurobiol 48(2):308–314. doi: 10.1007/s12035-013-8496-5 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Spampanato C, De Maria S, Sarnataro M, Giordano E, Zanfardino M, Baiano S, Carteni M, Morelli F (2012) Simvastatin inhibits cancer cell growth by inducing apoptosis correlated to activation of Bax and down-regulation of BCL-2 gene expression. Int J Oncol 40(4):935–941. doi: 10.3892/ijo.2011.1273 PubMedGoogle Scholar
  30. 30.
    Yu X, Pan Y, Ma H, Li W (2013) Simvastatin inhibits proliferation and induces apoptosis in human lung cancer cells. Oncol Res 20(8):351–357. doi: 10.3727/096504013X13657689382897 CrossRefPubMedGoogle Scholar
  31. 31.
    Johnson-Anuna LN, Eckert GP, Franke C, Igbavboa U, Muller WE, Wood WG (2007) Simvastatin protects neurons from cytotoxicity by up-regulating Bcl-2 mRNA and protein. J Neurochem 101(1):77–86. doi: 10.1111/j.1471-4159.2006.04375.x CrossRefPubMedGoogle Scholar
  32. 32.
    Franke C, Noldner M, Abdel-Kader R, Johnson-Anuna LN, Gibson Wood W, Muller WE, Eckert GP (2007) Bcl-2 upregulation and neuroprotection in guinea pig brain following chronic simvastatin treatment. Neurobiol Dis 25(2):438–445. doi: 10.1016/j.nbd.2006.10.004 CrossRefPubMedGoogle Scholar
  33. 33.
    Grewal SS, York RD, Stork PJ (1999) Extracellular-signal-regulated kinase signalling in neurons. Curr Opin Neurobiol 9(5):544–553. doi: 10.1016/S0959-4388(99)00010-0 CrossRefPubMedGoogle Scholar
  34. 34.
    Kim S, Lee KY, Koh SH, Park HH, Yu HJ, Lee YJ (2012) Role of the phosphatidylinositol 3-kinase and extracellular signal-regulated kinase pathways in the neuroprotective effects of cilnidipine against hypoxia in a primary culture of cortical neurons. Neurochem Int 61(7):1172–1182. doi: 10.1016/j.neuint.2012.08.010 CrossRefPubMedGoogle Scholar
  35. 35.
    Hetman M, Xia Z (2000) Signaling pathways mediating anti-apoptotic action of neurotrophins. Acta Neurobiol Exp 60(4):531–545Google Scholar
  36. 36.
    Fridovich I (1975) Superoxide dismutases. Annu Rev Biochem 44:147–159. doi: 10.1146/annurev.bi.44.070175.001051 CrossRefPubMedGoogle Scholar
  37. 37.
    Forsberg K, Andersen PM, Marklund SL, Brannstrom T (2011) Glial nuclear aggregates of superoxide dismutase-1 are regularly present in patients with amyotrophic lateral sclerosis. Acta Neuropathol 121(5):623–634. doi: 10.1007/s00401-011-0805-3 CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Barber SC, Mead RJ, Shaw PJ (2006) Oxidative stress in ALS: a mechanism of neurodegeneration and a therapeutic target. Biochim Biophys Acta 1762(11–12):1051–1067. doi: 10.1016/j.bbadis.2006.03.008 CrossRefPubMedGoogle Scholar
  39. 39.
    Forman HJ, Maiorino M, Ursini F (2010) Signaling functions of reactive oxygen species. Biochemistry 49(5):835–842. doi: 10.1021/bi9020378 CrossRefPubMedPubMedCentralGoogle Scholar
  40. 40.
    Stone JR, Yang S (2006) Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal 8(3–4):243–270. doi: 10.1089/ars.2006.8.243 CrossRefPubMedGoogle Scholar
  41. 41.
    Li Q, Spencer NY, Pantazis NJ, Engelhardt JF (2011) Alsin and SOD1(G93A) proteins regulate endosomal reactive oxygen species production by glial cells and proinflammatory pathways responsible for neurotoxicity. J Biol Chem 286(46):40151–40162. doi: 10.1074/jbc.M111.279711 CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Tanaka T, Tatsuno I, Uchida D, Moroo I, Morio H, Nakamura S, Noguchi Y, Yasuda T, Kitagawa M, Saito Y, Hirai A (2000) Geranylgeranyl-pyrophosphate, an isoprenoid of mevalonate cascade, is a critical compound for rat primary cultured cortical neurons to protect the cell death induced by 3-hydroxy-3-methylglutaryl-CoA reductase inhibition. J Neurosci 20(8):2852–2859PubMedGoogle Scholar
  43. 43.
    García-Román N, Alvarez AM, Toro MJ, Montes A, Lorenzo MJ (2001) Lovastatin induces apoptosis of spontaneously immortalized rat brain neuroblasts: involvement of nonsterol isoprenoid biosynthesis inhibition. Mol Cell Neurosci 17(2):329–341CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Seok-Ho Lee
    • 1
  • Na-Young Choi
    • 2
  • Hyun-Jeung Yu
    • 3
  • Jinse Park
    • 4
  • Hojin Choi
    • 1
  • Kyu-Yong Lee
    • 1
  • Yong-Min Huh
    • 5
  • Young Joo Lee
    • 1
  • Seong-Ho Koh
    • 1
    • 2
  1. 1.Department of NeurologyHanyang University College of MedicineGyeonggi-doSouth Korea
  2. 2.Department of Translational Medicine, Graduate School of Biomedical Science and EngineeringHanyang UniversitySeoulSouth Korea
  3. 3.Department of NeurologyBundang Jesaeng HospitalGwangjuSouth Korea
  4. 4.Department of Neurology, Haeundae Paik Hospital, College of MedicineInje UniversityBusanSouth Korea
  5. 5.Department of Radiology, College of MedicineYonsei UniversitySeoulSouth Korea

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