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Journal of Orthopaedic Surgery and Research
Published in: Journal of Orthopaedic Surgery and Research 1/2017

Open Access 01-12-2017 | Research article

Simvastatin protects ischemic spinal cord injury from cell death and cytotoxicity through decreasing oxidative stress: in vitro primary cultured rat spinal cord model under oxygen and glucose deprivation-reoxygenation conditions

Authors: Hye-Min Sohn, Jin-Young Hwang, Jung-Hee Ryu, Jinhee Kim, Seongjoo Park, Jin-woo Park, Sung-Hee Han

Published in: Journal of Orthopaedic Surgery and Research | Issue 1/2017

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Abstract

Background

Ischemia and the following reperfusion damage are critical mechanisms of spinal cord injury. Statins have been reported to decrease ischemia–reperfusion injury in many organs including the spinal cord. Anti-oxidative effect is one of the main protective mechanisms of statin against neuronal death and cytotoxicity. We hypothesized that statins’ anti-oxidative property would yield neuroprotective effects on spinal cord ischemia–reperfusion injury

Methods

Primary cultured spinal cord motor neurons were isolated from Sprague–Dawley rat fetuses. Ischemia–reperfusion injury model was induced by 60 min of oxygen and glucose deprivation (OGD) and 24 h of reoxygenation. Healthy and OGD cells were treated with simvastatin at concentrations of 0.1, 1, and 10 μM for 24 h. Cell viability was assessed using water-soluble tetrazolium salt (WST)-8, cytotoxicity with LDH, and production of free radicals with DCFDA (2′,7′-dichlorofluorescein diacetate).

Results

OGD reduced neuronal viability compared to normoxic control by 35.3%; however, 0.1–10 μM of simvastatin treatment following OGD improved cell survival. OGD increased LDH release up to 214%; however, simvastatin treatment attenuated its cytotoxicity at concentrations of 0.1–10 μM (p < 0.001 and p = 0.001). Simvastatin also reduced deteriorated morphological changes of motor neurons following OGD. Oxidative stress was reduced by simvastatin (0.1–10 μM) compared to untreated cells exposed to OGD (p < 0.001).

Conclusions

Simvastatin effectively reduced spinal cord neuronal death and cytotoxicity against ischemia–reperfusion injury, probably via modification of oxidative stress.
Literature
1.
Tator CH, Edmonds VE. Acute spinal cord injury: analysis of epidemiologic factors. Can J Surg. 1979;22:575–8.PubMed
2.
Celik M, Gokmen N, Erbayraktar S, Akhisaroglu M, Konakc S, Ulukus C, Genc S, Genc K, Sagiroglu E, Cerami A, Brines M. Erythropoietin prevents motor neuron apoptosis and neurologic disability in experimental spinal cord ischemic injury. Proc Natl Acad Sci U S A. 2002;99:2258–63.CrossRefPubMedPubMedCentral
3.
Sekhon LH, Fehlings MG. Epidemiology, demographics, and pathophysiology of acute spinal cord injury. Spine (Phila Pa 1976). 2001;26:S2–12.CrossRef
4.
McVicar JP, Luce JM. Management of spinal cord injury in the critical care setting. Crit Care Clin. 1986;2:747–58.PubMed
5.
Tator CH, Fehlings MG. Review of the secondary injury theory of acute spinal cord trauma with emphasis on vascular mechanisms. J Neurosurg. 1991;75:15–26.CrossRefPubMed
6.
Aki T, Toya S. Experimental study on changes of the spinal-evoked potential and circulatory dynamics following spinal cord compression and decompression. Spine (Phila Pa 1976). 1984;9:800–9.CrossRef
7.
Basaran R, Kaner T. C5 nerve root palsy following decompression of cervical spine with anterior versus posterior types of procedures in patients with cervical myelopathy. Eur Spine J. 2016;25:2050–9.CrossRefPubMed
8.
Madhavan K, Chieng LO, Foong H, Wang MY. Surgical outcomes of elderly patients with cervical spondylotic myelopathy: a meta-analysis of studies reporting on 2868 patients. Neurosurg Focus. 2016;40:E13.CrossRefPubMed
9.
Hasegawa K, Homma T, Chiba Y. Upper extremity palsy following cervical decompression surgery results from a transient spinal cord lesion. Spine (Phila Pa 1976). 2007;32:E197–202.CrossRef
10.
Vaughan CJ, Delanty N. Neuroprotective properties of statins in cerebral ischemia and stroke. Stroke. 1999;30:1969–73.CrossRefPubMed
11.
Jones SP, Trocha SD, Lefer DJ. Pretreatment with simvastatin attenuates myocardial dysfunction after ischemia and chronic reperfusion. Arterioscler Thromb Vasc Biol. 2001;21:2059–64.CrossRefPubMed
12.
Naidu BV, Woolley SM, Farivar AS, Thomas R, Fraga C, Mulligan MS. Simvastatin ameliorates injury in an experimental model of lung ischemia-reperfusion. J Thorac Cardiovasc Surg. 2003;126:482–9.CrossRefPubMed
13.
Sironi L, Cimino M, Guerrini U, Calvio AM, Lodetti B, Asdente M, Balduini W, Paoletti R, Tremoli E. Treatment with statins after induction of focal ischemia in rats reduces the extent of brain damage. Arterioscler Thromb Vasc Biol. 2003;23:322–7.CrossRefPubMed
14.
Devarajan P. Update on mechanisms of ischemic acute kidney injury. J Am Soc Nephrol. 2006;17:1503–20.CrossRefPubMed
15.
Sutherland BA, Minnerup J, Balami JS, Arba F, Buchan AM, Kleinschnitz C. Neuroprotection for ischaemic stroke: translation from the bench to the bedside. Int J Stroke. 2012;7:407–18.CrossRefPubMed
16.
Hayashi T, Rani PJ, Fukatsu A, Matsui-Hirai H, Osawa M, Miyazaki A, Tsunekawa T, Kano-Hayashi H, Iguchi A, Sumi D, Ignarro LJ. A new HMG-CoA reductase inhibitor, pitavastatin remarkably retards the progression of high cholesterol induced atherosclerosis in rabbits. Atherosclerosis. 2004;176:255–63.CrossRefPubMed
17.
Balduini W, Mazzoni E, Carloni S, De Simoni MG, Perego C, Sironi L, Cimino M. Prophylactic but not delayed administration of simvastatin protects against long-lasting cognitive and morphological consequences of neonatal hypoxic-ischemic brain injury, reduces interleukin-1beta and tumor necrosis factor-alpha mRNA induction, and does not affect endothelial nitric oxide synthase expression. Stroke. 2003;34:2007–12.CrossRefPubMed
18.
Cimino M, Gelosa P, Gianella A, Nobili E, Tremoli E, Sironi L. Statins: multiple mechanisms of action in the ischemic brain. Neuroscientist. 2007;13:208–13.CrossRefPubMed
19.
van der Most PJ, Dolga AM, Nijholt IM, Luiten PG, Eisel UL. Statins: mechanisms of neuroprotection. Prog Neurobiol. 2009;88:64–75.CrossRefPubMed
20.
Hayashi T, Hamakawa K, Nagotani S, Jin G, Li F, Deguchi K, Sehara Y, Zhang H, Nagano I, Shoji M, Abe K. HMG CoA reductase inhibitors reduce ischemic brain injury of Wistar rats through decreasing oxidative stress on neurons. Brain Res. 2005;1037:52–8.CrossRefPubMed
21.
Hong H, Zeng JS, Kreulen DL, Kaufman DI, Chen AF. Atorvastatin protects against cerebral infarction via inhibition of NADPH oxidase-derived superoxide in ischemic stroke. Am J Physiol Heart Circ Physiol. 2006;291:H2210–5.CrossRefPubMed
22.
Manzanero S, Santro T, Arumugam TV. Neuronal oxidative stress in acute ischemic stroke: sources and contribution to cell injury. Neurochem Int. 2013;62:712–8.CrossRefPubMed
23.
Pannu R, Barbosa E, Singh AK, Singh I. Attenuation of acute inflammatory response by atorvastatin after spinal cord injury in rats. J Neurosci Res. 2005;79:340–50.CrossRefPubMed
24.
Esposito E, Rinaldi B, Mazzon E, Donniacuo M, Impellizzeri D, Paterniti I, Capuano A, Bramanti P, Cuzzocrea S. Anti-inflammatory effect of simvastatin in an experimental model of spinal cord trauma: involvement of PPAR-alpha. J Neuroinflammation. 2012;9:81.PubMedPubMedCentral
25.
Anderson DK, Means ED, Waters TR, Spears CJ. Spinal cord energy metabolism following compression trauma to the feline spinal cord. J Neurosurg. 1980;53:375–80.CrossRefPubMed
26.
Hwang J, Han JI, Han S. Effect of pretreatment with simvastatin on spinal cord ischemia-reperfusion injury in rats. J Cardiothorac Vasc Anesth. 2013;27:79–85.CrossRefPubMed
27.
Saito T, Tsuchida M, Umehara S, Kohno T, Yamamoto H, Hayashi J. Reduction of spinal cord ischemia/reperfusion injury with simvastatin in rats. Anesth Analg. 2011;113:565–71.PubMed
28.
Lim JH, Lee JC, Lee YH, Choi IY, Oh YK, Kim HS, Park JS, Kim WK. Simvastatin prevents oxygen and glucose deprivation/reoxygenation-induced death of cortical neurons by reducing the production and toxicity of 4-hydroxy-2E-nonenal. J Neurochem. 2006;97:140–50.CrossRefPubMed
29.
Domoki F, Kis B, Gaspar T, Snipes JA, Parks JS, Bari F, Busija DW. Rosuvastatin induces delayed preconditioning against oxygen-glucose deprivation in cultured cortical neurons. Am J Physiol Cell Physiol. 2009;296:C97–105.CrossRefPubMed
30.
Jiang XY, Fu SL, Nie BM, Li Y, Lin L, Yin L, Wang YX, Lu PH, Xu XM. Methods for isolating highly-enriched embryonic spinal cord neurons: a comparison between enzymatic and mechanical dissociations. J Neurosci Methods. 2006;158:13–8.CrossRefPubMed
31.
Huang X, Sun J, Zhao T, Wu KW, Watanabe K, Xiao ZC, Zhu LL, Fan M. Loss of NB-3 aggravates cerebral ischemia by impairing neuron survival and neurite growth. Stroke. 2011;42:2910–6.CrossRefPubMed
32.
Brown M, Hart C, Tawadros T, Ramani V, Sangar V, Lau M, Clarke N. The differential effects of statins on the metastatic behaviour of prostate cancer. Br J Cancer. 2012;106:1689–96.CrossRefPubMedPubMedCentral
33.
Yamamoto Y, Mochida J, Sakai D, Nakai T, Nishimura K, Kawada H, Hotta T. Upregulation of the viability of nucleus pulposus cells by bone marrow-derived stromal cells: significance of direct cell-to-cell contact in coculture system. Spine (Phila Pa 1976). 2004;29:1508–14.CrossRef
34.
Deng X, Luan Q, Chen W, Wang Y, Wu M, Zhang H, Jiao Z. Nanosized zinc oxide particles induce neural stem cell apoptosis. Nanotechnology. 2009;20:115101.CrossRefPubMed
35.
Johnson-Anuna LN, Eckert GP, Franke C, Igbavboa U, Muller WE, Wood WG. Simvastatin protects neurons from cytotoxicity by up-regulating Bcl-2 mRNA and protein. J Neurochem. 2007;101:77–86.CrossRefPubMed
36.
Burd JF, Usategui-Gomez M. A colorimetric assay for serum lactate dehydrogenase. Clin Chim Acta. 1973;46:223–7.CrossRefPubMed
37.
Jayalakshmi K, Sairam M, Singh SB, Sharma SK, Ilavazhagan G, Banerjee PK. Neuroprotective effect of N-acetyl cysteine on hypoxia-induced oxidative stress in primary hippocampal culture. Brain Res. 2005;1046:97–104.CrossRefPubMed
38.
Amarenco P, Moskowitz MA. The dynamics of statins: from event prevention to neuroprotection. Stroke. 2006;37:294–6.CrossRefPubMed
39.
Liu L, Liu H, Yang F, Chen G, Zhou H, Tang M, Zhang R, Dong Q. Tissue kallikrein protects cortical neurons against hypoxia/reoxygenation injury via the ERK1/2 pathway. Biochem Biophys Res Commun. 2011;407:283–7.CrossRefPubMed
40.
Zacco A, Togo J, Spence K, Ellis A, Lloyd D, Furlong S, Piser T. 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors protect cortical neurons from excitotoxicity. J Neurosci. 2003;23:11104–11.PubMed
41.
Nagotani S, Hayashi T, Sato K, Zhang W, Deguchi K, Nagano I, Shoji M, Abe K. Reduction of cerebral infarction in stroke-prone spontaneously hypertensive rats by statins associated with amelioration of oxidative stress. Stroke. 2005;36:670–2.CrossRefPubMed
42.
Lathia JD, Chigurupati S, Thundyil J, Selvaraj PK, Mughal MR, Woodruff TM, Chan SL, Karamyan VT, Mattson MP, Arumugam TV. Pivotal role for beta-1 integrin in neurovascular remodelling after ischemic stroke. Exp Neurol. 2010;221:107–14.CrossRefPubMed
43.
Shabanzadeh AP, Shuaib A, Wang CX. Simvastatin reduced ischemic brain injury and perfusion deficits in an embolic model of stroke. Brain Res. 2005;1042:1–5.CrossRefPubMed
44.
Lefer AM, Scalia R, Lefer DJ. Vascular effects of HMG CoA-reductase inhibitors (statins) unrelated to cholesterol lowering: new concepts for cardiovascular disease. Cardiovasc Res. 2001;49:281–7.CrossRefPubMed
45.
Koh JY, Choi DW. Quantitative determination of glutamate mediated cortical neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Methods. 1987;20:83–90.CrossRefPubMed
46.
47.
Collard CD, Gelman S. Pathophysiology, clinical manifestations, and prevention of ischemia-reperfusion injury. Anesthesiology. 2001;94:1133–8.CrossRefPubMed
48.
Marttila RJ, Roytta M, Lorentz H, Rinne UK. Oxygen toxicity protecting enzymes in the human brain. J Neural Transm. 1988;74:87–95.CrossRefPubMed
49.
Makar TK, Nedergaard M, Preuss A, Gelbard AS, Perumal AS, Cooper AJ. Vitamin E, ascorbate, glutathione, glutathione disulfide, and enzymes of glutathione metabolism in cultures of chick astrocytes and neurons: evidence that astrocytes play an important role in antioxidative processes in the brain. J Neurochem. 1994;62:45–53.CrossRefPubMed
50.
Cafe C, Torri C, Bertorelli L, Tartara F, Tancioni F, Gaetani P, Rodriguez y Baena R, Marzatico F. Oxidative events in neuronal and glial cell-enriched fractions of rat cerebral cortex. Free Radic Biol Med. 1995;19:853–7.CrossRefPubMed
51.
Kontos HA, George E. Brown memorial lecture. Oxygen radicals in cerebral vascular injury. Circ Res. 1985;57:508–16.CrossRefPubMed
52.
Traystman RJ, Kirsch JR, Koehler RC. Oxygen radical mechanisms of brain injury following ischemia and reperfusion. J Appl Physiol (1985). 1991;71:1185–95.
53.
54.
Schouten O, Bax JJ, Dunkelgrun M, Feringa HH, van Urk H, Poldermans D. Statins for the prevention of perioperative cardiovascular complications in vascular surgery. J Vasc Surg. 2006;44:419–24.CrossRefPubMed
55.
Moulakakis KG, Matoussevitch V, Borgonio A, Gawenda M, Brunkwall J. Evidence that statins protect renal function during endovascular repair of AAAs. Eur J Vasc Endovasc Surg. 2010;40:608–15.CrossRefPubMed
56.
Guzik B, Sagan A, Ludew D, Mrowiecki W, Chwala M, Bujak-Gizycka B, Filip G, Grudzien G, Kapelak B, Zmudka K, Mrowiecki T, Sadowski J, Korbut R, Guzik TJ. Mechanisms of oxidative stress in human aortic aneurysms--association with clinical risk factors for atherosclerosis and disease severity. Int J Cardiol. 2013;168:2389–96.CrossRefPubMedPubMedCentral
57.
Karadimas SK, Gatzounis G, Fehlings MG. Pathobiology of cervical spondylotic myelopathy. Eur Spine J. 2015;24 Suppl 2:132–8.CrossRefPubMed
58.
Hu S, Dong HL, Li YZ, Luo ZJ, Sun L, Yang QZ, Yang LF, Xiong L. Effects of remote ischemic preconditioning on biochemical markers and neurologic outcomes in patients undergoing elective cervical decompression surgery: a prospective randomized controlled trial. J Neurosurg Anesthesiol. 2010;22:46–52.CrossRefPubMed
59.
Yaman O, Yaman B, Aydin F, Var A, Temiz C. Hyperbaric oxygen treatment in the experimental spinal cord injury model. Spine J. 2014;14:2184–94.CrossRefPubMed
60.
Abramov AY, Scorziello A, Duchen MR. Three distinct mechanisms generate oxygen free radicals in neurons and contribute to cell death during anoxia and reoxygenation. J Neurosci. 2007;27:1129–38.CrossRefPubMed
61.
Jung YS, Oh AY, Park HP, Hwang JW, Lim YJ, Jeon YT. Post-ischemic administration of pravastatin reduces neuronal injury by inhibiting Bax protein expression after transient forebrain ischemia in rats. Neurosci Lett. 2015;594:87–92.CrossRefPubMed
62.
Bosel J, Gandor F, Harms C, Synowitz M, Harms U, Djoufack PC, Megow D, Dirnagl U, Hortnagl H, Fink KB, Endres M. Neuroprotective effects of atorvastatin against glutamate-induced excitotoxicity in primary cortical neurones. J Neurochem. 2005;92:1386–98.CrossRefPubMed
63.
Loane DJ, Faden AI. Neuroprotection for traumatic brain injury: translational challenges and emerging therapeutic strategies. Trends Pharmacol Sci. 2010;31:596–604.CrossRefPubMedPubMedCentral
64.
Tanaka T, Tatsuno I, Uchida D, Moroo I, Morio H, Nakamura S, Noguchi Y, Yasuda T, Kitagawa M, Saito Y, Hirai A. 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. 2000;20:2852–9.PubMed
65.
Marz P, Otten U, Miserez AR. Statins induce differentiation and cell death in neurons and astroglia. Glia. 2007;55:1–12.CrossRefPubMed
66.
67.
Bu DX, Tarrio M, Grabie N, Zhang Y, Yamazaki H, Stavrakis G, Maganto-Garcia E, Pepper-Cunningham Z, Jarolim P, Aikawa M, Garcia-Cardena G, Lichtman AH. Statin-induced Kruppel-like factor 2 expression in human and mouse T cells reduces inflammatory and pathogenic responses. J Clin Invest. 2010;120:1961–70.CrossRefPubMedPubMedCentral
68.
Kureishi Y, Luo Z, Shiojima I, Bialik A, Fulton D, Lefer DJ, Sessa WC, Walsh K. The HMG-CoA reductase inhibitor simvastatin activates the protein kinase Akt and promotes angiogenesis in normocholesterolemic animals. Nat Med. 2000;6:1004–10.CrossRefPubMedPubMedCentral
69.
Ponce J, de la Ossa NP, Hurtado O, Millan M, Arenillas JF, Davalos A, Gasull T. Simvastatin reduces the association of NMDA receptors to lipid rafts: a cholesterol-mediated effect in neuroprotection. Stroke. 2008;39:1269–75.CrossRefPubMed
70.
Qi XF, Zheng L, Lee KJ, Kim DH, Kim CS, Cai DQ, Wu Z, Qin JW, Yu YH, Kim SK. HMG-CoA reductase inhibitors induce apoptosis of lymphoma cells by promoting ROS generation and regulating Akt, Erk and p38 signals via suppression of mevalonate pathway. Cell Death Dis. 2013;4:e518.CrossRefPubMedPubMedCentral
71.
Hsieh CH, Rau CS, Hsieh MW, Chen YC, Jeng SF, Lu TH, Chen SS. Simvastatin-induced heme oxygenase-1 increases apoptosis of Neuro 2A cells in response to glucose deprivation. Toxicol Sci. 2008;101:112–21.CrossRefPubMed
Metadata
Title
Simvastatin protects ischemic spinal cord injury from cell death and cytotoxicity through decreasing oxidative stress: in vitro primary cultured rat spinal cord model under oxygen and glucose deprivation-reoxygenation conditions
Authors
Hye-Min Sohn
Jin-Young Hwang
Jung-Hee Ryu
Jinhee Kim
Seongjoo Park
Jin-woo Park
Sung-Hee Han
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Journal of Orthopaedic Surgery and Research / Issue 1/2017
Electronic ISSN: 1749-799X
DOI
https://doi.org/10.1186/s13018-017-0536-9

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