Advanced Search
Korean J Pediatr 2012 July;55(7) :238-248.
Published online 2012 July 15.        doi:
Neuroprotective effects of L-carnitine against oxygenglucose deprivation in rat primary cortical neurons
Yu Jin Kim (Kim YJ)1, Soo Yoon Kim (Kim SY)2, Dong Kyung Sung (Sung DK)2, Yun Sil Chang (Chang YS)1, Won Soon Park (Park WS)1
1Department of Pediatrics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
2Samsung Biochemical Research Institute, Seoul, Korea
Corresponding Author: Yun Sil Chang ,Tel: +82-2-3410-3539, Fax: +82-2-3410-0043, Email:
Copyright © 2012 by The Korean Pediatric Society
Purpose: Hypoxic-ischemic encephalopathy is an important cause of neonatal mortality, as this brain injury disrupts normal mitochondrial respiratory activity. Carnitine plays an essential role in mitochondrial fatty acid transport and modulates excess acyl coenzyme A levels. In this study, we investigated whether treatment of primary cultures of rat cortical neurons with L-carnitine was able to prevent neurotoxicity resulting from oxygen-glucose deprivation (OGD). Methods: Cortical neurons were prepared from Sprague-Dawley rat embryos. L-Carnitine was applied to cultures just prior to OGD and subsequent reoxygenation. The numbers of cells that stained with acridine orange (AO) and propidium iodide (PI) were counted, and lactate dehydrogenase (LDH) activity and reactive oxygen species (ROS) levels were measured. The 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay and the terminal uridine deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assay were performed to evaluate the effect of L-carnitine (1 M, 10 M, and 100 M) on OGD-induced neurotoxicity. Results: Treatment of primary cultures of rat cortical neurons with L-carnitine significantly reduced cell necrosis and prevented apoptosis after OGD. L-Carnitine application significantly reduced the number of cells that died, as assessed by the PI/AO ratio, and also reduced ROS release in the OGD groups treated with 10 M and 100 M of L-carnitine compared with the untreated OGD group (P<0.05). The application of L-carnitine at 100 M significantly decreased cytotoxicity, LDH release, and inhibited apoptosis compared to the untreated OGD group (P<0.05). Conclusion: L-Carnitine has neuroprotective benefits against OGD in rat primary cortical neurons in vitro.
Keywords: L-carnitine | Neuroprotective effect | Oxygen-glucose deprivation | Neurons | Hypoxia-Ischemia
1. Johnston MV, Trescher WH, Ishida A, Nakajima W. Neurobiology of hypoxic-ischemic injury in the developing brain. Pediatr Res 2001;49:735–74
2. Vannucci RC. Hypoxic-ischemic encephalopathy. Am J Perinatol 2000;17:113–120.
3. Aliev G, Smith MA, Obrenovich ME, de la Torre JC, Perry G. Role of vascular hypoperfusion-induced oxidative stress and mitochondria failure in the pathogenesis of Azheimer disease. Neurotox Res 2003;5:491–504.
4. Gilland E, Puka-Sundvall M, Hillered L, Hagberg H. Mitochondrial function and energy metabolism after hypoxia-ischemia in the immature rat brain: involvement of NMDA-receptors. J Cereb Blood Flow Metab 1998;18:297–30
5. Gustavsson M, Anderson MF, Mallard C, Hagberg H. Hypoxic preconditioning confers long-term reduction of brain injury and improvement of neurological ability in immature rats. Pediatr Res 2005;57:305–309.
6. Wagner CL, Eicher DJ, Katikaneni LD, Barbosa E, Holden KR. The use of hypothermia: a role in the treatment of neonatal asphyxia?. Pediatr Neurol 1999;21:429–443.
7. Ma D, Hossain M, Pettet GK, Luo Y, Lim T, Akimov S, et al. Xenon preconditioning reduces brain damage from neonatal asphyxia in rats. J Cereb Blood Flow Metab 2006;26:199–208.
8. Clancy RR, McGaurn SA, Goin JE, Hirtz DG, Norwood WI, Gaynor JW, et al. Allopurinol neurocardiac protection trial in infants undergoing heart surgery using deep hypothermic circulatory arrest. Pediatrics 2001;108:61–70.
9. Asou H, Ono K, Uemura I, Sugawa M, Uyemura K. Axonal growth-related cell surface molecule, neurin-1, involved in neuron-glia interaction. J Neurosci Res 1996;45:571–587.
10. Himmelseher S, Pfenninger E, Georgieff M. Effects of basic fibroblast growth factor on hippocampal neurons after axonal injury. J Trauma 1997;42:659–664.
11. Koeppen AH, Dickson AC, McEvoy JA. The heterogeneous distribution of brain transferrin. J Neuropathol Exp Neurol 1995;54:395–403.
12. Houlgatte R, Mallat M, Brachet P, Prochiantz A. Secretion of nerve growth factor in cultures of glial cells and neurons derived from different regions of the mouse brain. J Neurosci Res 1989;24:143–152.
13. Rebouche CJ, Engel AG. Carnitine metabolism and deficiency syndromes. Mayo Clin Proc 1983;58:533–540.
14. Winter S, Jue K, Prochazka J, Francis P, Hamilton W, Linn L, et al. The role of L-carnitine in pediatric cardiomyopathy. J Child Neurol 1995;10 Suppl 2:S45–S51.
15. Fritz IB, Arrigoni-Martelli E. Sites of action of carnitine and its derivatives on the cardiovascular system: interactions with membranes. Trends Pharmacol Sci 1993;14:355–360.
16. Wainwright MS, Kohli R, Whitington PF, Chace DH. Carnitine treatment inhibits increases in cerebral carnitine esters and glutamate detected by mass spectrometry after hypoxia-ischemia in newborn rats. Stroke 2006;37:524–530.
17. Matsuishi T, Stumpf DA, Seliem M, Eguren LA, Chrislip K. Propionate mitochondrial toxicity in liver and skeletal muscle: acyl CoA levels. Biochem Med Metab Biol 1991;45:244–253.
18. Pande SV, Blanchaer MC. Reversible inhibition of mitochondrial adenosine diphosphate phosphorylation by long chain acyl coenzyme A esters. J Biol Chem 1971;246:402–411.
19. Loster H, Keller T, Grommisch J, Grunder W. Effects of L-carnitine and its acetyl and propionyl esters on ATP and PCr levels of isolated rat hearts perfused without fatty acids and investigated by means of 31P-NMR spectroscopy. Mol Cell Biochem 1999;200:93–102.
20. Wang D, Xia Y, Buja LM, McMillin JB. The liver isoform of carnitine palmitoyltransferase I is activated in neonatal rat cardiac myocytes by hypoxia. Mol Cell Biochem 1998;180:163–170.
21. Iijima T, Mishima T, Akagawa K, Iwao Y. Neuroprotective effect of propofol on necrosis and apoptosis following oxygen-glucose deprivation: relationship between mitochondrial membrane potential and mode of death. Brain Res 2006;1099:25–32.
22. Slivka A, Silbersweig D, Pulsinelli W. Carnitine treatment for stroke in rats. Stroke 1990;21:808–811.
23. Shenk JC, Liu J, Fischbach K, Xu K, Puchowicz M, Obrenovich ME, et al. The effect of acetyl-L-carnitine and R-alpha-lipoic acid treatment in ApoE4 mouse as a model of human Alzheimer's disease. J Neurol Sci 2009;283:199–206.
24. Zou X, Sadovova N, Patterson TA, Divine RL, Hotchkiss CE, Ali SF, et al. The effects of L-carnitine on the combination of, inhalation anesthetic-induced developmental, neuronal apoptosis in the rat frontal cortex. Neuroscience 2008;151:1053–1065.
25. Athanassakis I, Zarifi I, Evangeliou A, Vassiliadis S. L-carnitine accelerates the in vitro regeneration of neural network from adult murine brain cells. Brain Res 2002;932:70–78.
26. Loster H, Bohm U. L-carnitine reduces malondialdehyde concentrations in isolated rat hearts in dependence on perfusion conditions. Mol Cell Biochem 2001;217:83–90.
27. Long J, Gao F, Tong L, Cotman CW, Ames BN, Liu J. Mitochondrial decay in the brains of old rats: ameliorating effect of alpha-lipoic acid and acetyl-L-carnitine. Neurochem Res 2009;34:755–763.
28. Wainwright MS, Mannix MK, Brown J, Stumpf DA. L-carnitine reduces brain injury after hypoxia-ischemia in newborn rats. Pediatr Res 2003;54:688–695.
29. Hurtado O, Lizasoain I, Fernandez-Tome P, Alvarez-Barrientos A, Leza JC, Lorenzo P, et al. TACE/ADAM17-TNF-alpha pathway in rat cortical cultures after exposure to oxygen-glucose deprivation or glutamate. J Cereb Blood Flow Metab 2002;22:576–585.
30. Sauvageot CM, Stiles CD. Molecular mechanisms controlling cortical gliogenesis. Curr Opin Neurobiol 2002;12:244–249.
31. Banker GA. Trophic interactions between astroglial cells and hippocampal neurons in culture. Science 1980;209:809–810.
32. Mattson MP, Barger SW, Begley JG, Mark RJ. Calcium, free radicals, and excitotoxic neuronal death in primary cell culture. Methods Cell Biol 1995;46:187–216.
33. Brewer GJ, Torricelli JR, Evege EK, Price PJ. Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination. J Neurosci Res 1993;35:567–576.
34. Romera C, Hurtado O, Botella SH, Lizasoain I, Cardenas A, Fernandez-Tome P, et al. In vitro ischemic tolerance involves upregulation of glutamate transport partly mediated by the TACE/ADAM17-tumor necrosis factor-alpha pathway. J Neurosci 2004;24:1350–1357.
35. De Cristobal J, Cardenas A, Lizasoain I, Leza JC, Fernandez-Tome P, Lorenzo P, et al. Inhibition of glutamate release via recovery of ATP levels accounts for a neuroprotective effect of aspirin in rat cortical neurons exposed to oxygen-glucose deprivation. Stroke 2002;33:261–267.
36. Bank HL. Rapid assessment of islet viability with acridine orange and propidium iodide. In Vitro Cell Dev Biol 1988;24:266–273.
37. Meng XF, Zou XJ, Peng B, Shi J, Guan XM, Zhang C. Inhibition of ethanol-induced toxicity by tanshinone IIA in PC12 cells. Acta Pharmacol Sin 2006;27:659–664.
38. Gao F, Yi J, Yuan JQ, Shi GY, Tang XM. The cell cycle related apoptotic susceptibility to arsenic trioxide is associated with the level of reactive oxygen species. Cell Res 2004;14:81–85.
39. Gwag BJ, Lobner D, Koh JY, Wie MB, Choi DW. Blockade of glutamate receptors unmasks neuronal apoptosis after oxygen-glucose deprivation in vitro. Neuroscience 1995;68:615–619.
40. He F, Wu LX, Liu FY, Yang LJ, Zhang Y, Zhang HF, et al. Protection of hepatocyte growth factor on neurons subjected to oxygen-glucose deprivation/reperfusion. Sheng Li Xue Bao 2008;60:235–242.
41. Tastekin A, Gepdiremen A, Ors R, Buyukokuroglu ME, Halici Z. Protective effect of L-carnitine against bilirubin-induced neuronal cell death. Brain Dev 2006;28:436–439.
42. Grojean S, Koziel V, Vert P, Daval JL. Bilirubin induces apoptosis via activation of NMDA receptors in developing rat brain neurons. Exp Neurol 2000;166:334–341.
43. Onem G, Aral E, Enli Y, Oguz EO, Coskun E, Aybek H, et al. Neuroprotective effects of L-carnitine and vitamin E alone or in combination against ischemia-reperfusion injury in rats. J Surg Res 2006;131:124–130.
44. Burlina AP, Sershen H, Debler EA, Lajtha A. Uptake of acetyl-L-carnitine in the brain. Neurochem Res 1989;14:489–493.
45. Kim CS, Roe CR. Maternal and fetal tissue distribution of L-carnitine in pregnant mice: low accumulation in the brain. Fundam Appl Toxicol 1992;19:222–227.
46. Rebouche CJ. Carnitine function and requirements during the life cycle. FASEB J 1992;6:3379–3386.
47. Steiber AL, Weatherspoon LJ, Spry L, Davis AT. Serum carnitine concentrations correlated to clinical outcome parameters in chronic hemodialysis patients. Clin Nutr 2004;23:27–34.
48. De Vivo DC, Bohan TP, Coulter DL, Dreifuss FE, Greenwood RS, Nordli DR Jr, et al. L-carnitine supplementation in childhood epilepsy: current perspectives. Epilepsia 1998;39:1216–1225.
49. Pons R, De Vivo DC. Primary and secondary carnitine deficiency syndromes. J Child Neurol 1995;10 Suppl 2:S8–S24.
50. O'Donnell J, Finer NN, Rich W, Barshop BA, Barrington KJ. Role of L-carnitine in apnea of prematurity: a randomized, controlled trial. Pediatrics 2002;109:622–626.
PDF Links  PDF Links
Full text via DOI  Full text via DOI
  via Pubmed
Full text via PMC  Full text via PMC
via Pubreader  via PubReader
Download Citation  Download Citation
Supplementary Material  Supplementary Material
Neuroprotective effects of erythropoietin against hypoxic injury via modulation of the mitogen-activated protein kinase pathway and apoptosis  2017 June;60(6)
Neuroprotective effects of mild hypoxia in organotypic hippocampal slice cultures  2015 April;58(4)
Protective Effect of Probucol against Adriamycin-Induced Apoptosis in Cultured Rat Cardiac Myocytes  2000 June;43(6)
ERRATUM: Neurons by oxygen-glucose deprivation in organotypic hippocampal slice cultures  2008 December;51(12)
Register for e-submission
Register here to access the e-submission system of Korean J Pediatr for authors and reviewers.
Manuscript Submission
To submit a manuscript, please visit the Korean J Pediatr e-submission management system at, read the Instructions for Authors, and log into the Korean J Pediatr e-submission system. For assistance with manuscript submission, please contact:
Free archive
Anyone may access any past or current articles without logging in.
Korean Pediatric Society Office
#1606, Seocho World Officetel, 19 Seoun-ro, Seocho-gu, Seoul 137-070, Korea
TEL : +82-2-3473-7305    FAX : +82-2-3473-7307   E-mail:
BrowseCurrent IssueFor Authors and ReviewersAbout
Copyright© The Korean Pediatric Society. All right reserved.