Inhibition of Oxidative Stress-induced and Excitotoxic Neuronal Cell Damage by Xuesaitong Ruanjiaonang

혈색통연교낭(血塞通軟膠囊)의 산화적 및 흥분성 신경세포독성 억제작용

  • Cho Jungsook (Department of Pharmacology, College of Medicine, Dongguk University and Section of Neuroscience, Medical Institute of Dongguk University)
  • 조정숙 (동국대학교 의과대학 약리학교실 및 의학연구소 신경과학 연구부)
  • Published : 2005.02.01

Abstract

Xuesaitong Ruanjiaonang (XR), a soft capsule containing Panax notoginseng saponins as main ingredients, is believed to remove extravasated blood and increase cerebral blood flow by improving blood circulation, and therefore, has been used in China to treat ischemic stroke or hemiplegia caused by cerebral thrombosis. To characterize pharmacological actions of XR, the present study evaluated its effects on neuronal cell damage induced by various oxidative insults or excitotoxic amino acids in primary cultured rat cortical cells. The neuronal cell viability was not affected by XR with the exposure for 2 h at the concentrations tested in this study ($10{\sim}1000\;{\mu}g/ml$). However, significant reduction of the cell viability was observed when the cultured cells were exposed to XR at $1000\;{\mu}g/ml$ for 24 h. XR was found to concentration-dependently inhibit the oxidative neuronal damage induced by $H_{2}O_2$, xanthine/xanthine oxidase or $Fe^{2+}$/ascorbic acid. In addition, it dramatically inhibited the excitotoxic damage induced by glutamate or N-methyl-D-aspartate (NMDA). We found that the NMDA-induced neurotoxicity was inhibited more effectively and potently than the glutamate-induced toxicity. Moreover, XR was found to exert mild inhibition of lipid peroxidation induced by $Fe^{2+}$/ascorbic acid in rat brain homogenates and some 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity. Taken together, these results demonstrate neuroprotective and antioxidant effects of XR, showing inhibition of oxidative and excitotoxic damage in the cultured cortical neurons, as well as inhibition of lipid peroxidation and its radical scavenging activity. Considering that excitotoxicity and oxidative stress pl ay crucial roles in neuronal cell damage during ischemia and reperfusion, these results may provide pharmacological basis for its clinical usage to treat ischemic stroke.

Keywords

References

  1. Gan, F. Y. and Zhen, G. Z. : Chemical composition studies of Panax notoginseng. Chin. Pharm. J. 27, 138 (1992)
  2. Li, W. and Fitzloff, J. F. A. : Validated method for quantitative determination of saponins in notoginseng (Panax notoginseng) using high-performance liquid chromatography with evaporative light-scattering detection. J. Pharm. Pharmacol. 53, 1637 (2001) https://doi.org/10.1211/0022357011778241
  3. Ma, L., Xiao, P., Guo, B., Wu, J., Liang, F. and Dong, S. : Cerebral protective effects of compounds isolated from Traditional Chinese Herbs. Zhongguo Zhongyao Zazhi 24, 238 (1999)
  4. Wei, H., Zunping, Z., Jianxin, L., Heyang, Y., Jin, Z., Xianhua, H. and Fei, L. : Study on therapeutic window of opportunity for Panax notoginseng saponins following focal cerebral ischemia/reperfusion injury in rats. J. Chin. Med. Mat. 27, 25 (2004)
  5. Han, J.-A., Kuang, Y.-Q., Zhou, H.-T., Yang, L.-B., Zeng, F.-J., Sun, Z.-H., Hu, W.-Y. and Wang, X.-L. : Effects of Panax notoginseng saponins on malondialdehyde content of the brain following brain injury in rabbits. Zhongguo Bingli Shengli Zashi 16, 269 (2000)
  6. Ma, L., Xiao, P., Liang, F. and Wu, J. : Protective effects of panax notoginseng saponins on neurons of primary cortical cultures of rat. Zhongguo Yaoxue Zashi 33, 143 (1998)
  7. Cho, J., Joo, N. E., Kong, J.-Y., Jeong, D.-Y., Lee, K. D. and Kang, B.-S. : Inhibition of excitotoxic neuronal death by methanol extract of Acori graminei rhizoma in cultured rat cortical neurons. J. Ethnopharmacol. 73, 31 (2000) https://doi.org/10.1016/S0378-8741(00)00262-2
  8. Cho, J., Kong, J.-Y., Jeong, D.-Y., Lee, K. D., Lee, D. U. and Kang, B.-S. : NMDA receptor-mediated neuroprotection by essential oils from rhizomes of Acorus gramineus. Life Sci. 68, 1567 (2001) https://doi.org/10.1016/S0024-3205(01)00944-4
  9. Jung, Y.-S., Kang, T.-S., Yoon, J.-H., Joe, B.-Y., Lim, H.-J., Seong, C.-M., Park, W. K., Kong, J. Y., Cho, J. and Park, N. S. : Synthesis and evaluation of 4-hydroxyphenylacetic acid amides and 4-hydroxycinnamamides as antioxidants. Bioorg. Med. Chem. Lett. 12, 2599 (2002) https://doi.org/10.1016/S0960-894X(02)00479-1
  10. Dok-Go, H., Lee, K. H., Kim, H. J. Lee, E. H., Lee, J., Song, Y. S., Lee, Y. H., Jin, C., Lee, Y. S. and Cho, J. : Neuroprotective effects of antioxidative flavonoids, quercetin, (+)-dihydroquercetin and quercetin 3-methyl ether, isolated from Opuntia ficus-indica var. saboten, Brain Res. 965, 130 (2003) https://doi.org/10.1016/S0006-8993(02)04150-1
  11. Cho, J., Kim, Y. H., Kong, J.-Y., Yang, C.-H. and Park, C.-G. : Protection of cultured rat cortical neurons from excitotoxicity by asarone, a major essential oil component in the rhizomes of Acorus gramineus. Life Sci. 71, 591 (2002) https://doi.org/10.1016/S0024-3205(02)01729-0
  12. 조정숙, 양재하, 박창국, 이희순, 김영호 : 뇌졸중 치료 생약 추출물의 흥분성 신경독성 억제효과. 약학회지 44, 29 (2000)
  13. Hansen, M. B., Nielsen, S. E. and 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 (1989) https://doi.org/10.1016/0022-1759(89)90397-9
  14. Cho, J. and Lee, H.-K. : Wogonin inhibits excitotoxic and oxidative neuronal damage in primary cultured rat cortical cells. Eur. J. Pharmacol. 485, 105 (2004) https://doi.org/10.1016/j.ejphar.2003.11.064
  15. Xie, C., Lovell, M. A., Xiong, S., Kindy, M. S., Guo, J.-T., Xie, J., Amaranth, V., Montine, T. J. and Markesbery, W. R. : Expression of glutathione-S-transferase isozymes in the SY5Y neuroblastoma cell line increases resistance to oxidative stress. Free Radic. Biol. Med. 31, 73 (2001) https://doi.org/10.1016/S0891-5849(01)00553-6
  16. 중화인민공화국약전. 제1부. 중화인민공화국약전위원회편 (1990)
  17. Reiter, R. J. : Oxidative processes and antioxidative defense mechanism in the aging brain. FASEB J. 9, 526 (1995) https://doi.org/10.1096/fasebj.9.7.7737461
  18. Sauer, D. and Fagg, G. E. : Excitatory amino acids, excitotoxicity and neurodegenerative disorders. In: Krogsgaard-Larsen, P., Hansen, J. J., eds. Excitatory amino acid receptors. Ellis Horwood, London, 13 (1992)
  19. Bigge, C. F. and Malone, T. C. : Agonists, antagonists and modulators of the N-methyl-D-aspartic acid (NMDA) and $\alpha$-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) subtypes of glutamate receptors. Curr. Opin. Ther. Patents July, 951 (1993)
  20. Lynch, D. R. and Guttmann, R. P. : Excitotoxicity: Perspectives based on N-methyl-D-aspartate receptor subtypes. J. Pharmacol. Exp. Ther. 300, 717 (2002) https://doi.org/10.1124/jpet.300.3.717
  21. Halliwell, B. : Reactive oxygen species and the central nervous system. J. Neurochem. 59, 1609 (1992) https://doi.org/10.1111/j.1471-4159.1992.tb10990.x
  22. Ma, L.-Y., Xiao, P.-G., Liang, F.-Q., Chi, M.-G. and Dong, S.-J. : Effect of saponins of Panax notoginseng on synaptosomal $^{45}Ca$ uptake. Zhongguo Yaoli Xuebao 18, 213 (1997)
  23. Ma, L. and Xiao, P. : Effects of Panax notoginseng saponins on synaptosomal glutamate release and its specific binding to glutamic receptor. Zhongguo Yaolixue Tongbao 14, 311 (1998)
  24. Xuejiang, W., Ichikawa, H. and Konishi, T. : Antioxidant potential of Qizhu Tang, a Chinese herbal medicine, and the effect on the cerebral oxidative damage after ischemia reperfusion in rats. Biol. Pharm. Bull. 24, 558 (2001) https://doi.org/10.1248/bpb.24.1
  25. Ng, T. B., Liu, F. and Wang, H. X. : The antioxidant effects of aqueous and organic extracts of Panax quinquefolium, Panax notoginseng, Codonopsis pilosula, Pseudostellaria heterophylla and Glehnia littoralis. J. Ethnopharmacol. 93, 285 (2004) https://doi.org/10.1016/j.jep.2004.03.040