Mapping of Rat Hippocampal Neurons With NeuN After Ischemia/Reperfusion and Ginkgo Biloba Extract (EGb 761) Pretreatment

Springer Science and Business Media LLC - Tập 26 - Trang 1191-1202 - 2006
Iveta Domoráková1, Jozef Burda2, Eva Mechírová1, Marianna Feriková1
1Department of Histology and Embryology, Faculty of Medicine, P. J. Šafárik University, Košice, Slovak Republic
2Institute of Neurobiology, Slovak Academy of Sciences, Košice, Slovak Republic

Tóm tắt

1. The neuroprotective effect of Ginkgo biloba extract (EGb 761) against transient forebrain ischemia following 7 days of reperfusion was studied in male Wistar rats after four-vessel occlusion for 20 min. 2. NeuN, a neuronal specific nuclear protein was used for immunohistochemical detection of surviving pyramidal neurons in the hippocampus, as well as counterstaining with hematoxylin in the same sections for detection of neurons that underwent delayed neuronal death and for glial nuclei staining. GFAP immunohistochemistry was used for detection of astrocytes in the studied area of CA1 region. 3. In the group of rats pretreated 7 days with Ginkgo biloba extract (EGb 761), following 20 min of ischemia and 7 days of reperfusion without EGb 761, increased number of NeuN immunoreactive cells were counted in the most vulnerable CA1 pyramidal layer of hippocampus. On the other hand, the group of rats with 7 days of EGb 761 pretreatment following 20 min of ischemia and 7 days of reperfusion with EGb 761 showed decreased number of surviving NeuN immunoreactive CA1 pyramidal cells in comparison with the first above-mentioned experimental group. 4. Increased number of reactive astrocytes immunolabeled for GFAP (Glial fibrilary acidic protein) was observed in both experimental groups in the stratum oriens and stratum lacunosum and moleculare. 5. Twenty minutes of ischemia is lethal for most population of CA1 pyramidal cell layer. Our results showed that prophylactic oral administration of Ginkgo biloba extract (EGb 761) in the dose 40 mg/kg/day during the 7 days protects the most vulnerable CA1 pyramidal cells against 20 min of ischemia.

Tài liệu tham khảo

Bodsch, W., Takahashi, K., Barbier, A., Ophoff, B. G., and Hossmann, K. A. (1985). Cerebral protein synthesis and ischemia. Prog. Brain Res. 63:197–210. Burda, J., Martin, M. E., Garcia, A., Alcazar, A., Fando, J. L., and Salinas, M. (1994). Phosphorylation of the alpha subunit of initiation factor 2 correlates with the inhibition of translation following transient cerebral ischemia in the rat. Biochem. J. 302:335–338. Calapai, G., Crupi, A., Firenzuoli, F., Marciano, M. C., Squadrito, F., Inferrera, G., Parisi, G., Rizzo, A., Crisafulli, C., Fiore, A., and Caputi, A. P. (2000). Neuroprotective effect of Ginkgo biloba extract in brain ischemia are mediated by inhibition of nitric oxide synthesis. Life Sci. 67:2673–2683. Chandrasekaran, K., Mehrabian, Z., Spinnewyn, B., Drieu, K., and Fiskum, G. (2001). Neuroprotective effects of bilobalide, a component of the Ginkgo biloba extract (EGb 761), in gerbil global brain ischemia. Brain Res. 922:282–292. Chen, Y., and Swanson, R. A. (2003). Astrocytes and brain injury. J. Cereb. Blood Flow Metab. 23:137–149. Clark, W. M., Rinker, L. G., Lessov, N. S., Lowery, S. L., and Cipolla, M. J. (2001). Efficacy of antioxidant therapies in transient focal ischemia in mice. Stroke 32:1000–1004. D'Ambrosio, A. L., Pinsky, D. J., and Connolly, E. S. (2001). The role of a complement cascade in ischemia/reperfusion injury: Implication for neuroprotection. Mol. Med. 7:367–382. DeFeudis, F. V. (1998). Ginkgo biloba extract (EGb 761), Ullstein Medical, Wiesbaden, pp. 1–401. Drieu, K. (1986). Preparation and definition of Ginkgo biloba extract. Presse Med. 15:1455–1457. Dykens, J. A. (1994). Isolated cerebral and cerebellar mitochondria produce free radicals when exposed to elevated CA2+ and Na+: Implications of neurodegeneration. J. Neurochem. 63:584–591. Feriková, M., Mechírová, E., and Domoráková, I. (2004). Demonstration of ubiquitin following ischemia and reperfusion of spinal cord neurons in the rabbit. Acta Vet. Brno 73:51–54. Fiskum, G., Murphy, A. N., and Beal, M. F. (1999). Mitochondria in neurodegeneration: Acute ischemia and chronic neurodegenerative diseases. J. Cereb. Blood Flow Metab. 19:351–369. Gdovinová, Z. (2001).Tanakan v liečbe neurologických ochorení. Slov. Lekár 11:12–15. Hrehorovská, M., Burda, J., Domoráková, I., and Mechírová, E. (2004). Effect of Tanakan on postischemic activity of protein synthesis machinery in the rat brain. Gen. Physiol. Biophys. 23:457–465. Joyeux, M., Lobstein, A., Anton, A., and Mortier, F. (1995). Comparative antilipid peroxidant, antinecrotic and scavenging properties of terpenes and biflavones from Ginkgo biloba. Planta Med. 61:126–129. Kim, Y. S., Pyo, M. K., Park, K. M., Park, P. H., Hahn, B. S., Wu, S. J., and Yun-Choi, H. S. (1998). Antiplatelet and antithrombotic effects of a combination of ticlopidine and Ginkgo biloba extract (EGb 761). Thromb. Res. 91:33–38. Kirino, T. (1982). Delayed neuronal death in the gerbil hippocampus following ischemia. Brain Res. 239:57–69. Leis, J., Bekar, L., and Walz, W. (2005). Potassium homeostasis in the ischemic brain. Glia 50:407–416. Lo, E. H., Dalkara, T., and Moskowitz, M. A. (2003). Mechanisms, challenges and opportunities in stroke. Nat. Rev. Neurosci. 4:399–415. Marcocci, L., Packer, L., Droy-Lefaix, M. T., Sekaki, A., and Gardes-Albert, M. (1994). Antioxidant action of Ginkgo biloba extract EGb 761. Methods Enzymol. 234:462–475. Matthias, K., Kirchhoff, F., Seifert, G., Huttmann, K., Matyash, M., Kettenmann, H., and Steinhauser, C. (2003). Segregated expression of AMPA-type glutamate receptors and glutamate transporters defines distinct astrocyte population in the mouse hippocampus. J. Neurosci. 23:1750–1758. Mechírová, E., and Domoráková, I. (2002). NADPH-diaphorase activity in the spinal cord after ischemic injury and the effects of pretreatment with Ginkgo biloba extract (EGb 761). Acta Histochem. 104(4):427–430. Morin, D., Hauet, T., Spedding, M., and Tillement, J. P. (2001). Mitochondria as a target for antiischemic drugs. Adv. Drug Deliv. Rev. 49:151–174. Nishino, K., and Nowak, T. S. Jr. (2004). Time course and cellular distribution of hsp27 and hsp72 stress protein expression in a quantitative gerbil model of ischemic injury and tolerance: Thresholds for hsp72 induction and hilar lesioning in the contex of ischemic preconditioning. J. Cereb. Blood Flow Metab. 24:167–178. Oyama, Y., Chikahisa, L., Ueha, T., Kanemaru, K., and Noda, K. (1996). Ginkgo biloba extract protects brain neurons against oxidative stress induced by hydrogen peroxide. Brain Res. 712:349–352. Pascual, J. M., Carceller, F., Roda, J. M., and Cerdan, S. (1998). Glutamate, glutamine, and GABA as substrates for the neuronal and glial compartments after focal cerebral ischemia in rats. Stroke 29:1048–1056. Peng, H., Li, Y. F., and Sun, S. G. (2003). Effects of Ginkgo biloba extract on acute cerebral ischemia in rats analyzed by magnetic resonance spectroscopy. Acta Pharmacol. Sin. 24:467–471. Petito, C. K., and Babiak, T. (1982). Early proliferative changes in astrocytes in postischemic noninfarcted rat brain. Ann. Neurol. 11:510–518. Petito, C. K., Feldmann, E., Pulsinelli, W. A., and Plum, F. (1987). Delayed hippocampal damage in humans following cardiorespiratory arrest. Neurology 37:1281–1286. Pietri, S., Maurelli, E., Drieu, K., and Culcasi, M. (1997). Cardioprotective and anti- oxidant effects of the terpenoid constituents of Ginkgo biloba extract (EGb 761). J. Mol. Cell. Cardiol. 29:733–742. Puisieux, F., Deplanque, D., Bulckaen, H., Maboudou, P., Gelé, P., Lhermitte, M., Lebuffe, G., and Bordet, R. (2004). Brain ischemic preconditioning is abolished by antioxidant drugs but does not up-regulate superoxide dismutase and glutathion peroxidase. Brain Res. 1027:30–37. Pulsinelli, W. A. (1985). Selective neuronal vulnerability: Morphological and molecular characteristics. Prog. Brain Res. 63:29–37. Pulsinelli, W. A., and Brierley, J. B. (1979). A new model of bilateral hemispheric ischemia in the unanesthetized rat. Stroke 10:267–272. Pulsinelli, W. A., Brierley, J. B., and Plum, F. (1982a). Temporal profile of neuronal damage in a model of transient forebrain ischemia. Ann. Neurol. 11:491–498. Pulsinelli, W. A., Levy, D. E., and Duffy, T. E. (1982b). Regional cerebral blood flow and glucose metabolism following transient forebrain ischemia. Ann. Neurol. 11:499–509. Roy, S., Kobushi, H., Sen, C. K., Gohil, K., Droix-Lefaix, M. T., and Packer, L. (1998). Ginkgo biloba extract (EGb 761): Antioxidant properties and regulation of gene expression. In Packer, L., and Christen, Y. (eds.), Advances in Ginkgo Biloba Extract Research, Ginkgo biloba Extract (EGb 761): Lessons from Cell Biology, Vol. 7, Elsevier, Paris, pp. 1–17. Schmidt-Kastner, R., Paschen, W., Ophoff, B. G., and Hossmann, K. A. (1989). A modified four vessel occlusion model for inducing incomplete forebrain ischemia in rats. Stroke 20:938–946. Seif-El-Nasr, M., and El-Fattah, A. A. (1995). Lipid peroxide, phospholipids, glutathione levels and superoxid dismutase activity in rat brain after ischemia: Effects of Ginkgo biloba extract. Pharmacol. Res. 32:273–278. Shen, J. G., and Zhou, D. Y. (1995). Efficiency of Ginkgo biloba extract (EGb 761) in antioxidant protection against myocardial ischemia and reperfusion injury. Biochem. Mol. Biol. Int. 35:125–134. Spinnewyn, B., Blavet, N., and Clostre, F. (1986). Effects of Ginkgo biloba extract on a cerebral ischemia model in gerbils. Presse Méd. 15:1511–1515. Swanson, R. A., Ying, W., and Kauppinen, T. M. (2004). Astrocyte influences on ischemic neuronal death. Curr. Mol. Med. 4:193–205. Westman, J., Drieu, K., and Sharma, H. S. (2000). Antioxidant compounds EGb-761 and BN-520 21 attenuate heat shock protein (HSP 72 kD) response, edema and cell changes following hyperthermic brain injury. An experimental study using immunohistochemistry in the rat. Amino Acids 19:339–350. Zhang, W. R., Hayashi, T., Kitagawa, H., Sasaki, C., Sasaki, K., Warita, H., Wang, J. M., Shiro, Y., Uchida, M., and Abe, K. (2000). Protective effect of Ginkgo extract on rat brain with transient middle cerebral artery occlusion. Neurol. Res. 22:517–521.