Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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The Red-ginseng Extract Alters the Cell Cycle and Viability in the Human Neuronal Stem Cells

홍삼추출액의 인간성체신경줄기세포 증식과 세포사 관련 세포주기의 변화에 대한 효과

  • Kim, Hyun-Jung (Department of Neurology, Seoul National University Hospital) ;
  • Kang, La-Mi (Department of Neurology, Seoul National University Hospital) ;
  • Ahn, Jin-Young (Department of Neurology, Gangnam General Hospital Public Corporation) ;
  • Han, Jung-Soon (Department of Nutrition and Food Sciences of Junior College of Allied Health Sciences of Korea University/Diet Research Institute of InJae University) ;
  • Kim, Seung-U. (Brain Disease Research Center, Ajou University) ;
  • Lee, Kwang-Woo (Department of Neurology, Seoul National University Hospital) ;
  • Kim, Man-Ho (Department of Neurology, Seoul National University Hospital)
  • 김현정 (서울대학교 의과대학 신경과) ;
  • 강라미 (서울대학교 의과대학 신경과) ;
  • 안진영 (지방공사 강남병원 신경과) ;
  • 한정순 (고려대학교 보건대학 식품영양학과/인제대학교부설 다이어트연구소) ;
  • 김승업 (아주대학교 뇌질환연구소) ;
  • 이광우 (서울대학교 의과대학 신경과) ;
  • 김만호 (서울대학교 의과대학 신경과)
  • Published : 2004.03.01
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Abstract

The present study is to determine whether the Red-ginseng extract has a proliferative or cytotoxic effect on the human neuronal stem cells(hNSCs). The hNSCs were grown and incubated with different doses of Red-ginseng extract. We tested the proliferative or cytotoxic effects by MTT and FACS analysis. Cell viability cell cycle analysis, DNA fragmentation, and bax or PARP expressions were evaluated. The hNSCs showed a proliferafe trend with its peak concentration at 0.3 $\mu\textrm{g}$/$m\ell$. Beyond this point, higher doses decreased viabilities and showed a cytotoxic effect at 10 $\mu\textrm{g}$/$m\ell$. There was a tendency of increased S and G2/M phases during cell proliferation. In a cytotoxic condition, decreased S phase and increased G0/G1 phases were noted, suggesting cell cycle arrest. The cytotoxic effect was associated with increase DNA fragmentation in a dose-dependent manner, However PARP cleavage or bax expression was not detected. Our results suggest that Red-ginseng extract has dual effects, the cell proliferative or cytotoxic effect, on hNSCs in vitro with dose-dependent manner.

홍삼추출액의 신경성체줄기세포 성장과 생존에 미치는 영향을 분석하고자 human neuronal stem cell line인 F3 cell을 배양한 후 홍삼추출액을 여러 농도로 희석하여 MTT assay로 cell viability를 측정하였고 FACS analysis로 cell Cycle변화를 측정하였다. 특정한 농도에서는 세포가 증식되는 경향을 보였으며 농도가 증가되면서 viability가 감소되는 현상을 확인할 수 있었다. Cell cycle 분석상 세포증식시에는 5 phase 및 G2/M phase가 증가되는 경향을 보였고, viability가 감소되면서 S phase가 감소되고 G0/G1 phase 가 증가 되었다. 한편 DNA fragmentation이 cell viability감소에 따라 증가되었으나, Caspase 3 activation 또는 Bax expression과는 관련성이 적었다.

Keywords

References

  1. Chen, J., Li, Y., Wang, L., Lu, M., Zhang, X. and Chopp, M. : Therapeutic benefit of intracerebral transplantation of bone marrow stromal cells after cerebral ischemia in rats. J Neurol sci. 189, 49-57 (2001) https://doi.org/10.1016/S0022-510X(01)00557-3
  2. Chen, J., Li, Y., Wang, L., Zhang, Z., Lu, D. and Lu, M. et al. : Therapeutic benefit of intravenous administration of bone marrow stromal cells afrer cerebral ischemia in rats. Stroke. 32, 1005-1011 (2001) https://doi.org/10.1161/01.STR.32.4.1005
  3. Li, Y., Chen, J., Wang, L., Lu, M. and Chopp, M. : Treatment of stroke in rat with intracarotid administration of marrow stromal cells. Neurology. 56, 1666-167 (2001) https://doi.org/10.1212/WNL.56.12.1666
  4. Zhao, L.R., Duan, W.M., Reyes, M., Keene, C.D., Verfaillie, C.M. and Low, W.C. : Human bone marrow stem cells exhibit neural phenotypes and ameliorate neurological deficits after grafting into the ischemic brain of rats. Exp Neurol. 174, 11-20 (2002) https://doi.org/10.1006/exnr.2001.7853
  5. Chen, J., Sanberg, P.R., Li, Y., Wang, L., Lu, M. and Willing, A.E. et al. : Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats. Stroke. 32, 2682-2688 (2001) https://doi.org/10.1161/hs1101.098367
  6. Arvidsson, A., Collin, T., Kirik, D., Kokaia, Z. and Lindvall, O. : Neuronal replacement from endogenous precursors in the adult brain after stroke. Nat Med. 8, 963-970 (2002) https://doi.org/10.1038/nm747
  7. Takahashi, J., Palmer, T.D. and Gage, F.H. : Retinoic acid and neurotrophins collaborate to regulate neurogenesis in adult-derived neural stem cell cultures. J Neurobiol. 38, 65-81 (1999) https://doi.org/10.1002/(SICI)1097-4695(199901)38:1<65::AID-NEU5>3.0.CO;2-Q
  8. Li, Y., Chen, J., Chen, X.G., Wang, L., Gautam, S. C. and Xu, Y. X. et al. : Human marrow stromal cell therapy for stroke in rat: neurotrophins and functional recovery. Neurology. 59, 514-523 (2002)
  9. Jiang, F., DeSilva, S. and Turnbull, J. : Beneficial effect of ginseng root in SOD-1 (G93A) transgenic mice. J Neurol Sci. 180, 52-54 (2000) https://doi.org/10.1016/S0022-510X(00)00421-4
  10. Hahn, J., Nah, S.Y., Nah, J.J., Uhm, D.Y. and Chung, S. : Ginsenosides inhibit capsaicin-activated channel in rat sensory neurons. Neurosci Lett. 287, 45-48 (2000) https://doi.org/10.1016/S0304-3940(00)01119-8
  11. Kim, Y.C., Kim, S. R., Markelonis, G. J. and Oh, T. H. : Ginsenosides Rb1 and Rg3 protect cultured rat cortical cells from glutamate-induced neurodegeneration. J Neurosci Res. 53, 426-432 (1998) https://doi.org/10.1002/(SICI)1097-4547(19980815)53:4<426::AID-JNR4>3.0.CO;2-8
  12. Lim, J. H., Wen, T. C., Matsuda, S., Tanaka, J., Maeda, N., Peng, H., Aburaya, J., Ishihara, K. and Sakanaka, M. : Protection of ischemic hippocampal neurons by ginsenoside Rb1, a main ingredient of ginseng root. Neurosci Res. 28, 191-200 (1997) https://doi.org/10.1016/S0168-0102(97)00041-2
  13. Wen, T. C., Yoshimura, H., Matsuda, S., Lim, J. H. and Sakanaka, M. : Ginseng root prevents learning disability and neuronal loss in gerbils with 5-minute forebrain ischemia. Acta Neuropathol (Berl). 91, 15-22 (1996) https://doi.org/10.1007/s004010050387
  14. Liu, M. and Zhang, J. T. : Protective effects of ginsenoside Rb1 and Rg1 on cultured hippocampal neurons. Yao Xue Xue Bao. 30, 674-678 (1995)
  15. Himi, T., Saito, H. and Nishiyama, N. : Effect of ginseng saponins on the survival of cerebral cortex neurons in cell cultures. Chem Pharm Bull (Tokyo). 37, 481-484 (1989) https://doi.org/10.1248/cpb.37.481
  16. Flax, J. D., Aurora, S., Yang, C., Simonn, C., Wills, A. N. and Billinghurst, L. L. et al. : Engraftable human neural stem cells respond to developmental cues, replace neurons, and express foreign genes. Nat Biotechnol. 16, 1033-1039 (1998) https://doi.org/10.1038/3473
  17. Ourednik, V., Ourednik, J., Flax, J.D., Zawada, W.M., Hutt, C. and Yang, C. et al. : Segregation of human neural stem cells in the developing primate forebrain. Science. 293, 1820-1824 (2001) https://doi.org/10.1126/science.1060580
  18. Vekrellis, K., McCarthy, M. J., Watson, A., Whitfield, J., Rubin, L. L. and Ham, J. : Bax promotes neuronal cell death and is downregulated during the development of the nervous system. Development. 124, 1239-1249 (1997)
  19. Morrison, R. S., Kinoshita, Y., Johnson, M. D., Chatan, S., Ho, J. T. and Garden, G. : Neuronal survival and cell death signaling pathways. Adv Exp Med Biol. 513, 41-86(2002)
  20. Mongan, P. D., Karaian, J., Van Der Schuur,, B. M., Via, D. K. and Sharma, P. : Pyruvate prevents poly-ADP rivose polymerase (PARP) activation, oxidative damage, and pyru-vate dehydrogenase deactivation during hemorrhagic shock in swine. J. Surg Res. 112, 180-188 (2003) https://doi.org/10.1016/S0022-4804(03)00148-3
  21. Weise, J., Isenmann, S. and Bahr, M. : Increased expression and activation of poly (ADP-ribose) polymerase (PARP) contribute to retinal ganglion cell death following rat optic nerve transection, Cell Death Differ. 8, 801-807(2001) https://doi.org/10.1038/sj.cdd.4400872
  22. Lee, Y. W., Ha, M. S. and Kim, Y. K. : $H_2O_2$-induced cell death in human glioma cells: role of lipid peroxidation and PARP activation. Neurochem Res. 26, 337-343 (2001) https://doi.org/10.1023/A:1010993428770