Inhibition Effect of Zizania latifolia on Apoptosis Induced by $H_2O_2$ in Neuro2A Cell

$H_2O_2$로 유발된 Neuro2A 신경세포고사에 대한 줄풀의 억제 효과

  • Park, Won-Hyung (Department of Oriental Rehabilitation Medicine, Collage of Oriental Medicine, Sangji University) ;
  • Cha, Yun-Yeop (Department of Oriental Rehabilitation Medicine, Collage of Oriental Medicine, Sangji University)
  • 박원형 (상지대학교 한의과대학 한방재활의학과) ;
  • 차윤엽 (상지대학교 한의과대학 한방재활의학과)
  • Published : 2005.08.01

Abstract

The purpose of this study was to examine the inhibition effect of Zizania latifolia that has been used heart disease, Diabetes Mellitus and Skin disease for a long time on apoptosis induced by $H_2O_2$ in Neuro2A cell. Neuro2A cells were cultivated in RPMI(GibcoBRL) with $5\%$, FBS and treated with $H_2O_2$, and Zizania latifolia. We measured the cell viability and analyzed DNA fragmentation. Activity of PARP, Cytochrome C, caspase-9, caspase-3, p53, p21, Bax and Bcl-2 in the cell was examined by using western blot. The cell viability in Zizania latifolia treatment (60ug/ml<) decreased significantly compared with that of none treatment. (p<0.001) Zizania latifolia increased cell viability about twice as much as that being injury by $H_2O_2$. (Zizania Latifolia 20ug/ml, $H_2O_2$ 200uM, P<0.001) DNA fragmentation developed by $H_2O_2$, but was not developed in Zizania latifolia treatment. PARP, Cytochrome C, caspase-9 and caspase-3 activated all by $H_2O_2$ but were not activated in Zizania latifolia treatment.. P53, P2l and Bu activated by $H_2O_2$, and Bcl-2 got into inactivation. But the opposite results appeared in Zizania latifolia treatment. In conclusion, these results suggest that Zizania latifolia inhibit the development of DNA fragmentation and apoptosis by $H_2O_2$ and the antioxidant action of Zizania latifolia is effective. More researches about effect of Zizania latifolia are considered to need.

Keywords

References

  1. Cutler, R.G. Antioxidants, aging and longevity. Free Radicals in Biology, Academic Press. 6, 371-424, 1988.
  2. Feher, J., Cosmos, G., Vereckei, A. The free radical theory of aging. Free Radicals Reactions in Medicine. Berlin, Springer-Verlag. pp 57-59, 1987.
  3. Halliwell, B., Gutteridge, J.M. The chemistry of oxygen radicals and other oxygen-derived species. In free radicals in biology and medicine. 2nd Ed. Oxford, Clarendon Press. pp 22-31, 1989.
  4. Halliwell, B., Gutteridge, J.M. Protection against oxidants in biological systems : the superoxide theory of oxygen toxicity. In free radicals in biology and medicine. 2nd Ed. Oxford, Clarendon Press. pp 86-89, 1989.
  5. Hertog, M.C. Hollman, P.C. Potential health effects of the dietary flavonid quercetin. Eur J Clin Nutr 50(2):63-66, 1996.
  6. Benavente-Garcisa, O., J. C. Marrin, F.R. Ortuno, A. Rio. Uses and properties of citurs flavonoids, J. Agric. Food Chem 45, 4505-4515, 1997.
  7. 정성제, 이진희, 송효남, 성낙술, 이승은, 백남인. 약용 식물 추출물의 항산화 활성 검색. 한국응용생명화학회지 47(1):135-140, 2004.
  8. 김은영, 백인희, 김정현, 김성란, 류미라. 항산화활성을 나타내는 약용식물 소재 탐색. 한국식품과학회지 36(2):333-338, 2004.
  9. 尹國炳, 張俊根. 몸에 좋은 山野草. 서울, 石悟出版社. p 410, 1989.
  10. 中藥大辭典. 서울, 醫聖堂. p 2052, 1994
  11. 동식, 김근하, 김영호, 최승문. 고려치료경험 내과편. 의학과학출판사. p 71, 76, 114, 122, 132, 166, 345, 386, 1996.
  12. Pryor, W. A Free radical in biology. Involvement of radical reaction in aging and carcinogenesis in medicinal chemistry. Elservier, Amsterdam. pp 331-361, 1997.
  13. Simon, R.H., Scogging, C.M., Patterson, D. Hydrogen peroxide cause the fatal injury to human fibroblast exposed to oxygen radicals. J. Biol. Chem 266, 7181-7186, 1981.
  14. Bokov, A., Chaudhuri, A., Richardson, A. The role of oxidative damage and stress in aging. Mech Aging Dev 125(10-11):811-826, 2004.
  15. Harman, D. Free radical theory of aging. Mutation Res 275(3-6):257-266, 1992.
  16. Husain, S.R, Cillard, J., Cillard, P. Hydroxy radical scavenging activity of flavonoids. Phytochemistry 26(9):2489-2491, 1987.
  17. Chandra, J., Samali, A, Orrenius, S. Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 29(3-4):323-333, 2000.
  18. Kerr, J.F., Wyllie, A.H., Currie, A.R. Apoptosis a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26(4):239-257, 1972.
  19. Li, P., Nijhawan, D., Budihardjo, I., Srinivasula, S.M., Ahmad, M., Alnemri, E.S., et al. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91(4):479-489, 1977.
  20. Soldani, C., Scovassi, A.I. Poly(ADP-ribose) polymerase-1 cleavage during apoptosis. an update. Apoptosis 7(4):321-328, 2002.
  21. Bert, T., Kenneth, W.K. P53 function and dysfunction. Cell 70, 523-526, 1995.
  22. Lane, D.P. A death in the life of P53. Nature 362(6423): 849-852, 1993.
  23. 안원근 노화과정에 있어서 P21 유전자의 역할. 생화학분자생물학뉴스. 한국생화학분자생물학회 23(2):80-81, 2003.
  24. Yang, E., Korsmeyer, S.K. Molecular thanatosis: A discourse on the bcl-2 family and cell death. Blood 88(2):386-401, 1996.
  25. Oltvai, Z., Milliman, C, Korsmeyer, S. J. bcl-2 heterodimerizes in vivo with a conserved homolog, Bax that accelerates programmed cell death. Cell 74(4):609-619, 1993.
  26. 고경석. 가을에 꽃 피는 야생식물. 서울 일진사. p 56, 2004.
  27. 李時珍. 本草網目. 北京, 人民衛生出版社. p 1366, 1982.
  28. 許浚. 對譯東醫寶鑑. 서울, 법인문화사. p 1956, 1999.
  29. 李埏. 編注 醫學入門. 서울, 醫聖堂. p 331, 1994.
  30. 冠宗奭. 本草衍義. 서울, 醫聖堂. p 74, 1994.
  31. 최진규. 약이 되는 우리풀 . 꽃, 나무 2. 서울, 한문화 p 141-147, 2003.
  32. 과학.백과사전출판사 편. 약초의 성분과 이용. 서울, 일월서각. p 652, 1994.
  33. 김복영, 김규식, 박영대 축산폐수의 오염물질제거를 위한 수초선발이용연구. 한국환경농학회지 7(2):111-116, 1988.
  34. 김하송, 임병선. 농경지 배출수의 수질개선을 위한 수생식물의 정화능과 활용방안에 관한 연구. 환경관리학회지 4(2):1-8, 1988
  35. Liu, Z., Wang, Y., Shen, Y., Guo, W., Hao, S., Liu, B. Extensive alterations in DNA methylation and transcription in rice caused by introgression from Zizania latifolia. Plant Mol Biol 54(4):571-582, 2004.
  36. Ryu, S.N., Park, S.Z., Kim, H.Y. Antioxidant Activity in Rice Cultivar, Wild Rice, and Barley. Korean J. Crop Sci 47(1):54-61, 2002.
  37. 박상원, 송춘호. 瓦松藥鍼液이 腎臟細胞에서 $H_2O_2$에 의한 細胞死亡 및 DNA 損傷에 미치는 影響. 대한침구학회지 18(1):88-99, 2001.
  38. 안성훈, 구성태, 김선영, 김경식, 손인철. $H_2O_2$로 유발된 뇌신경세포 상해에 대한 구진의 보호효과. 대한침구학회지 21(3):29-41, 2004.