DOI QR코드

DOI QR Code

Glutamate가 유도하는 세포독성으로부터 신경세포를 보호하는 상백피 추출물의 효과

Neuroprotective Effect of Extracts from Root Bark of Morus alba on Glutamate-induced Cytotoxicity in Neuronal Cells.

  • 김현정 (경남대학교 식품생명학과) ;
  • 김지현 (경남대학교 식품생명학과) ;
  • 손은순 (경남대학교 식품생명학과) ;
  • 이정민 (경남대학교 식품생명학과) ;
  • 박해룡 (경남대학교 식품생명학과)
  • Kim, Hyun-Jung (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Kim, Ji-Hyun (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Son, Eun-Soon (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Lee, Jeung-Min (Department of Food Science and Biotechnology, Kyungnam University) ;
  • Park, Hae-Ryong (Department of Food Science and Biotechnology, Kyungnam University)
  • 발행 : 2009.07.30

초록

본 연구에서는 glutamate로 유도된 세포독성으로부터 신경세포를 보호하는 상백피(Morus alba) 추출물의 활성을 확인하기 위하여 N18-RE-105 세포주를 이용하여 MTT reduction assay, LDH release assay 및 광학 현미경을 이용하여 형태학적인 변화를 관찰하였다. 그 결과, 상백피 methanol 추출물에서 농도 의존적으로 신경세포 보호효과가 나타났으며, 50 본 연구에서는 glutamate 로 유도된 세포독성으로부터 신경세포를 보호하는 상백피 (Morus alba) 추출물의 활성을 확인하기 위하여 NI8-RE-I05 세포주를 이용하여 MTT reduction assay, LDH release assay 및 광학 현 미 경 을 이 용하여 형태학적인 변화를 관찰하였다. 그 결과y 상백피 methanol 추출물에서 농도 의존적으로 신경세포 보호효과가 나타났으며, 50 ${\mu}g$/ml 농도에서는 80% 이상의 세포생존율을 확인할수 있었다. 이 결과는 N18-RE-105 세포주의 LDH release assay와 형태학적 변화에서도 일치하는 결과를 확인하였다. 가장 높은 활성을 보인 상백피 methanol 추출물을 hexane, diethyl ether, ethyl acetate, water 층으로 분획하여 각 각 1, 5, 10 ${\mu}g$/ml 농도로 처리 시 hexane 층에서 48.0%, 65.6%, 71.5%로 가장 높은 신경세포 보호효과를 확인할 수 있었다. 따라서 상백피 추출물은 glutamate 에 의한 세포독성으로부터 신경세포 손상을 억제하며 신경세포를 보호하는 효과가 있다는 것을 알 수 있었다.

This study evaluated the neuroprotective effect of extracts from the root bark of Morus alba (MA) against glutamate-induced cytotoxicity in neuronal cells. Glutamate-induced cytotoxicity was shown by MTT reduction assay. The neuroprotective effects of methanol, ethanol, and acetone extracts from MA against glutamate-induced cytotoxicity were measured. Among the three extracts, the methanolic extracts showed the highest protective effect, as determined by the results of an morphological assay, a lactate dehydrogenase release assay. Furthermore, the methanol extracts were fractionated sequentially with hexane, diethyl ether, ethyl acetate, and water layer according to degree of polarity. The hexane fractions exhibited a neuroprotective effect against glutamate-stressed N18-RE-105 cells. Therefore, these results suggest that extracts of MA could be a new potential candidate as a protective substance against glutamate-induced cytotoxicity.

키워드

참고문헌

  1. Chen, L. and L. Y. Hunag. 1991. Sustained potentiation of NMDA receptor-mediated glutamate responses through activation of protein kinase C by a mu opioid. Neuron 7, 319-326 https://doi.org/10.1016/0896-6273(91)90270-A
  2. Cowburn, R., J. Hardy, P. Roverts, and R. Briggs. 1988. Regional distribution of pre- and postsynaptic glutamatergic function in Alzheimer’s disease. Brain Res. 452, 403-407 https://doi.org/10.1016/0006-8993(88)90048-0
  3. Coyle, J. T. and P. Puttfarecken. 1993. Oxidative stress, glutamate, and neurodegenerative disorders. Science 262, 689-695 https://doi.org/10.1126/science.7901908
  4. Choi, B. W., E. M. Hur, J. H. Lee, D. J. Jun, and K. T. Kim. 2005. Protein kinase C$\delta$-mediated proteasomal degradation of MAP kinase phosphatase-1 contributes of glutamate-induced neuronal cell death. J. Cell Sci. 119, 1329-1340 https://doi.org/10.1242/jcs.02837
  5. Dillon-Carter, O., C. Conejero, M. Poltorak, and W. J. Freed. 1998. N18-RE-105 cells: differentiation and activation of p53 in response to glutamate and adriamycin is blocked by SV40 large T antigen tsA58. Cell Tissue Res. 291, 191-205 https://doi.org/10.1007/s004410050990
  6. D'Orlando, C., M. R. Celio, and B. Schwaller. 2002. Calretinin and calbindin D-28k, but not parvalbumin protect against glutamate-induced delayed excitotoxicity in trasfected N18-RE 105 neuroblastoma-retina hybrid cells. Brain Res. 945, 181-190 https://doi.org/10.1016/S0006-8993(02)02753-1
  7. Duffy, S., A. So, and T. H. Murphy. 1998. Activation of endogenous antioxidant defenses in neuronal cells prevents free radical-madiated damage. J. Neurochem. 71, 69-77 https://doi.org/10.1046/j.1471-4159.1998.71010069.x
  8. Fonnum, F. 1984. Glutamate: a neurotransmitter in mammalian brain. J. Neurochem. 42, 1-11 https://doi.org/10.1111/j.1471-4159.1984.tb09689.x
  9. Hara, J., D. Gerashchenko, J. P. Wisor, T. Sakurai, X. Xie, and T. S. Kilduff. 2009. Thyrotropin-releasing hormone increases behavioral arousal through modulation of hypocretin/orexin neurons. J. neruosci. 29, 3705-3714 https://doi.org/10.1523/JNEUROSCI.0431-09.2009
  10. Harman, D. 1981. The aging process. P. Natl. Acad. Sci. USA 78, 7124-7128 https://doi.org/10.1073/pnas.78.11.7124
  11. Hikino, H., T. Mizuno, Y. Oshima, and C. Konno. 1985. Isolation and hypoglycemic activity of Moran A, a glycoprotein of Mousalba root barks. Planta Med. 51, 159-160 https://doi.org/10.1055/s-2007-969435
  12. Jin, D. Q., C. S. Lim, J. K. Hwang, I. H. Ha, and J. S. Han. 2005. Anti-oxidant and anti-inflammatory activity of macelignan in murine hippocampal cell line and primary culture of rat microglial cells. Biochem. Biophy. Res. Co. 331, 1264-1269 https://doi.org/10.1016/j.bbrc.2005.04.036
  13. Kang, T. H., H. R. Oh, S. M. Jung, J. H. Ryu, M. W. Park, Y. K. Park, and S. Y. Kim. 2006. Enhancement of neuroprotection of mulberry leaves (Morus alba L.) prepared by the anaerobic treatment against ischemic damage. Biol. Rharm. Bull. 29, 270-274 https://doi.org/10.1248/bpb.29.270
  14. Kim, M. J., S. J. Choi, H. K. Kim, C. J. Kim, B. Hong, and Y. J. Kim. 2007. Activation effects of allium tuberosum Rottl. on choline acetyltrasnferase. Biosci. Biotech. Bioch. 71, 226-230 https://doi.org/10.1271/bbb.60314
  15. Kim, J. Y., J. H. Hwang, M. R. Cha, B. D. Choi, S. U. Choi, H. P. Park, and Y. I. Hwnag. 2006. Growth-inhibitory effects of the plocamium telfairiae extracts on cancer cells. Jorunal of Life Science 4, 659-663
  16. Kim, S. Y., H. S. Lee, K. S. Ryu, E. J. Lee, and Y. C. Kim. 1999. protective effets of extracts of Mori Cortex Radicis on corbon tetrachloride-induced hepatotoxicity. Yakhak Hoeji. 43, 391-396
  17. Lee, M. R., C. S. Han, D. Y. Han, E. Park, S. C. Lee, and H. R. Park. 2007. Protective effect of neuronal cell on glutamate- induced oxidative stress from viola mandshurica extracts. J. Korean Soc. Appl. Biol. Chem. 51, 79-83
  18. Lim, C. S., D. Q. Jin, J. Y. Sung, J. H Lee, H. G. Choi, I. H. Ha, and J. S. Han. 2006. Antioxidant and anti-inflammatory activities of the nethanolic extract of neorhodomela aculeate in hippocampal and microglial cells. Biol. Pharm. Bull. 29, 1212-1216 https://doi.org/10.1248/bpb.29.1212
  19. Monaghan, D. T., R. J. Bridges, and C. W. Cotman. 1989. The excitatory amino acid receptors: their classes, pharmacology and distinct properties in the function of the central nervous system. Annu. Rev. Pharmacol. 29, 365-402 https://doi.org/10.1146/annurev.pa.29.040189.002053
  20. Noh, H. S., Y. S. Hah, R. Nilufar, J. Han, J. H. Bong, S. S. Kang, G. J. Cho, and W. S. Choi. 2006. Acetoacetate protects neuronal cells from oxidative glutamate toxicity. J. Neurosic. Res. 83, 702-709 https://doi.org/10.1002/jnr.20736
  21. Nomura, T., T. Fukai, S. Yamada, and M. Katayanagi. 1976. Phenolic constituents of the cultivated Mulberry tree (Morus alba L.). Chem. Pharm. Bull. 24, 2898-2900 https://doi.org/10.1248/cpb.24.2898
  22. Olanwo, C. W. and W. G. Tatton. 1999. Etiology and pathogenesis of Parkinson’s disease. Annu. Rev. Neurosic. 22, 123-144 https://doi.org/10.1146/annurev.neuro.22.1.123
  23. Parfenova, H., S. Basuroy, S. Bhattacharya, D. Tcheranova, Y. Qu, R. F. Regan, and C. W. Leffler. 2005. Glutamate induces oxidative stress and apoptosis in cerebral vascular endothelial cells: contributions of HO-1 and HO-2 to cytoprotection. Am. J. Physiol. Cell Physiol. 290, 1399-1401 https://doi.org/10.1152/ajpcell.00386.2005
  24. Weiloch, T. 1985. Hypoglycemai-induced neuronal damage prevented by an N-methyl-D-aspartate antagonists. Science 230, 681-683 https://doi.org/10.1126/science.2996146
  25. Yoon, M. Y., J. Y. Kim, J. H. Hwang, M. R. Cha, M. R. Lee, K. J. Jo, and H. R. park. 2007. Protective effect of methanolic extracts from Dendrobium nobile Lindl. On H2O2-induced neurotoxicity in PC12 cells. J. Korean soc. Appl. Chem. 50, 63-67
  26. Zhang, W., F. Han, J. He, and C. Duan. 2008. HPLC-DAD-ESI-MS/MS analysis and antioxidant activities of nonanthocyanin phenolics in mulberry (Morus alba L.). J. Food Sci. 73, C512-518 https://doi.org/10.1111/j.1750-3841.2008.00854.x