Neuroprotective Effects of Hydroxyfullerene in Rats Subjected to Global Cerebral Ischemia

  • Kim, Young-Ock (Ginseng and Medicinal Plants Research Institute, Rural Development Administration) ;
  • Kim, Hak-Jae (Kohwang Medical Research Institute, School of Medicine, Kyung Hee University) ;
  • Kim, Su-Kang (Kohwang Medical Research Institute, School of Medicine, Kyung Hee University) ;
  • Yoon, Bum-Chul (Department of Physical Therapy, College of Health Science, Korea University)
  • Published : 2008.09.30

Abstract

Oxidative stress is believed to contribute to the neuronal damage induced by cerebral ischemia/reperfusion injury. The present study was undertaken to evaluate the possible antioxidant neuroprotective effect of hydroxyfullerene (a radical absorbing cage molecule) against neuronal death in hippocampal CA1 neurons following transient global cerebral ischemia in the rat. Transient global cerebral ischemia was induced in male Wistar rats by four vessel- occlusion (4VO) for 10 min. Lipid peroxidation in brain tissues was determined by measuring the concentrations of thiobarbituric acid-reactive substances (TBARS). Furthermore, the apoptotic effects of ${H_2}{O_2}$ on PC12 cells were also investigated. Cell viabilities were measured using MTT [3-(4,5-dimethylthiazolyl-2)-2,-5-diphenyltetrazolium bromide] assays. Hydroxyfullerene, when administered to rats at 0.3-3 mg/kg i.p. at 0 and 90 minutes after 4-VO was found to significantly reduce CA1 neuron death by 72.4% on hippocampal CA1 neurons. Our findings suggest that hydroxyfullerene protects neurons from transient global cerebral injury in the rat hippocampus by reducing oxidative stress and lipid peroxidation levels, which contribute to apoptotic cell death.

References

  1. Krusic, P. J. et al. Radical reactions of C60. Science 254:1183-1185 (1991) https://doi.org/10.1126/science.254.5035.1183
  2. Bosi, S., Da Ros, T., Spalluto, G. & Prato, M. Fullerene derivatives: an attractive tool for biological applications. Eur J Med Chem 38:913-923 (2003) https://doi.org/10.1016/j.ejmech.2003.09.005
  3. Dugan, L. L. et al. Buckminsterfullerenol free radical scavengers reduce excitotoxic and apoptotic death of cultured cortical neurons. Neurobiol Dis 3:129-135 (1996) https://doi.org/10.1006/nbdi.1996.0013
  4. Kirino, T. Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res 239:57-69 (1982) https://doi.org/10.1016/0006-8993(82)90833-2
  5. Graham, S. H. & Chen, J. Programmed cell death in cerebral ischemia. J Cereb Blood Flow Metab 21:99-109 (2001) https://doi.org/10.1097/00004647-200102000-00001
  6. Wang, Q. et al. Neuroprotective mechanisms of curcumin against cerebral ischemia-induced neuronal apoptosis and behavioral deficits. J Neurosci Res 82: 138-148 (2005) https://doi.org/10.1002/jnr.20610
  7. Wilson, J. X. & Gelb, A. W. Free radicals, antioxidants, and neurologic injury: possible relationship to cerebral protection by anesthetics. J Neurosurg Anesthesiol 14:66-79 (2002) https://doi.org/10.1097/00008506-200201000-00014
  8. Chan, P. H. Role of oxidants in ischemic brain damage. Stroke 27:1124-1129 (1996) https://doi.org/10.1161/01.STR.27.6.1124
  9. Dugan, L. L. et al. Fullerene-based antioxidants and neurodegenerative disorders. Parkinsonism Relat Disord 7:243-246 (2001) https://doi.org/10.1016/S1353-8020(00)00064-X
  10. Huang, S. S. et al. Neuroprotective effect of hexasulfobutylated C60 on rats subjected to focal cerebral ischemia. Free Radic Biol Med 30:643-649 (2001) https://doi.org/10.1016/S0891-5849(00)00505-0
  11. Esterbauer, H., Schaur, R. J. & Zollner, H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med 11:81-128 (1991) https://doi.org/10.1016/0891-5849(91)90192-6
  12. Fiers, W., Beyaert, R., Declercq, W. & Vandenabeele, P. More than one way to die: apoptosis, necrosis and reactive oxygen damage. Oncogene 18:7719-7730 (1999) https://doi.org/10.1038/sj.onc.1203249
  13. Nitatori, T. et al. Delayed neuronal death in the CA1 pyramidal cell layer of the gerbil hippocampus following transient ischemia is apoptosis. J Neurosci 15: 1001-1011 (1995) https://doi.org/10.1523/JNEUROSCI.15-02-01001.1995
  14. Mason, R. B. et al. Production of reactive oxygen species after reperfusion in vitro and in vivo: protective effect of nitric oxide. J Neurosurg 93:99-107 (2000) https://doi.org/10.3171/jns.2000.93.1.0099
  15. Collino, M. et al. Oxidative stress and inflammatory response evoked by transient cerebral ischemia/reperfusion: effects of the PPAR-alpha agonist WY14643. Free Radic Biol Med 41:579-589 (2006) https://doi.org/10.1016/j.freeradbiomed.2006.04.030
  16. Chan, P. H. Reactive oxygen radicals in signaling and damage in the ischemic brain. J Cereb Blood Flow Metab 21:2-14 (2001) https://doi.org/10.1097/00004647-200101000-00002
  17. Chueh, S. C. et al. Decrease of free radical level in organ perfusate by a novel water-soluble carbon-sixty, hexa (sulfobutyl)fullerenes. Transplant Proc 31:1976 -1977 (1999)
  18. Dugan, L. L. et al. Carboxyfullerenes as neuroprotective agents. Proc Natl Acad Sci U S A 94:9434-9439 (1997)
  19. Cagle, D. W. et al. In vivo studies of fullerene-based materials using endohedral metallofullerene radiotracers. Proc Natl Acad Sci U S A 96:5182-5187 (1999)
  20. Jin, H. et al. Polyhydroxylated C(60), fullerenols, as glutamate receptor antagonists and neuroprotective agents. J Neurosci Res 62:600-607 (2000) https://doi.org/10.1002/1097-4547(20001115)62:4<600::AID-JNR15>3.0.CO;2-F
  21. Hansen, M. B., Nielsen, S. E. & Berg, K. Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 119:203-210 (1989) https://doi.org/10.1016/0022-1759(89)90397-9
  22. Pulsinelli, W. A. & Brierley, J. B. A new model of bilateral hemispheric ischemia in the unanesthetized rat. Stroke 10:267-272 (1979) https://doi.org/10.1161/01.STR.10.3.267
  23. Lee, H. et al. Flavonoid wogonin from medicinal herb is neuroprotective by inhibiting inflammatory activation of microglia. Faseb J 17:1943-1944 (2003) https://doi.org/10.1096/fj.03-0057fje
  24. Zhang, D. L., Zhang, Y. T., Yin, J. J. & Zhao, B. L. Oral administration of Crataegus flavonoids protects against ischemia/reperfusion brain damage in gerbils. J Neurochem 90:211-219 (2004) https://doi.org/10.1111/j.1471-4159.2004.02480.x
  25. Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254 (1976) https://doi.org/10.1016/0003-2697(76)90527-3