Inhibition Effect on Neuro2A Cell by Apoptosis of Zizania latifolia Rhizoma

줄풀 줄기의 Neuro2A 신경세포고사에 대한 억제 효과

  • Cha Yun-Yeop (Department of Oriental Rehabilitation Medicine, College of Oriental Medicine, Sangji University)
  • 차윤엽 (상지대학교 한의과대학 한방재활의학과)
  • Published : 2006.02.01

Abstract

To prevent human body injury from oxidative stress, antioxidants are very important and many research about antioxidants are generally being conducted. Hydrogen peroxide($H_2O_2$) that is one of vitality oxygen species has been seen that cause various diseases, DNA damage and gene change. The purpose of this study was to examine the inhibition effect of Zizania latifolia Rhizoma 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 Rhizoma. 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 dy using western blot. The results obtained were as Follows: The cell viability in Zizania latifolia Rhizoma treatment (60ug/ml<) decreased significantly compared with that of none treatment. (P<0.001) Zizania latifolia Rhizoma increased cell viability about twice as much as that being injury by $H_2O_2$. (Zizania Latifolia Rhizoma 20ug/ml, $H_2O_2$ 200uM, P<0.001) DNA fragmentation developed by $H_2O_2$, but was not developed in Zizania latifolia Rhizoma treatment. PARP, Cytochrome C, caspase-9 and caspase-3 activated all by $H_2O_2$ but were not activated in Zizania latifolia Rhizoma treatment. P53, P2l and Bax activated dy $H_2O_2$, and Bcl-2 got into inactivation. But the opposite results appeared in Zizania latifolia Rhizoma treatment. In conclusion, these results suggest that Zizania latifolia Rhizoma inhibit the development of DNA fragmentation and apoptosis by $H_2O_2$ and the antioxidant action of Zizania latifolia Rhizoma is effective. More researches about effect of Zizania latifolia Rhizoma are considered to need.

Keywords

References

  1. 최진호. 노화의 메카니즘과 연구방향. 생화학뉴스. 한국생화학회 5(3):39-53, 1985
  2. 沈吉浩. 노화의 개념과 예방. 성문출판사. pp 15-17, 1987
  3. Cohen, G. The generation of hydroxyl radicals in biological system. Photobiol. 28, 669-675, 1978 https://doi.org/10.1111/j.1751-1097.1978.tb06993.x
  4. 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
  5. 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
  6. Hertog, M.G.L., P.C.H. Hollman. Potential health effects of the dietary flavonid quercetin. Eur J Clin Nutr 50(2):63-66, 1996
  7. 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 https://doi.org/10.1021/jf970373s
  8. 中藥大辭典. 서울, 醫聖堂. p 2052, 1994
  9. 동식, 김근하, 김영호, 최승문. 고려치료경험 내과편. 의학과학출판사. p 71, 76, 114, 122, 132, 166, 345, 386, 1996
  10. 박원형. H2O2로 유발된 Neuro2A 신경세포고사에 대한 줄풀의 억제 효과. 상지대학교 석사학위논문. 2005
  11. 두호경 편저. 동의신계학. 서울, 동양의학연구원. pp 1327-1329, 1993
  12. 김숙희, 김화영. 老化, 서울, 민음사, pp 77-85, 94, 1991
  13. Cutler, R.G. Antioxidant, aging and longevity. Free Radicals in Biology(ed. Pryor, W.), Acedimic Press. 6, 371-424, 1984
  14. Feher, J., Cosmos, G., Vereckei, A. The free radical theory of aging Free Radicals Reactions in Medicine. Springer-Verlag. Berlin. pp 57-59, 1987
  15. 김영곤, 김영균. 프리라디칼. 서울, 여문각. pp 31-35, 98-101, 259-260, 278-286, 396-400, 425-426, 564-568, 1997
  16. 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 https://doi.org/10.1038/bjc.1972.33
  17. Sarraf, C.E., Bowen, I.D. Proportions of mitotic and apoptotic cells in a range of untreated experimental tumours. Cell Tissue Kinet 21(1):45-49, 1988
  18. 홍원식 편. 精校黃帝內經. 서울, 東洋醫學硏究院. pp 19-20, 11, 246, 301, 1981
  19. 南京中醫學院醫經敎硏組. 黃帝內經素問譯解. 上海, 上海科學技術出版社. pp 4-5, 1983
  20. 南京中醫學院醫經敎硏組. 黃帝內經素問譯解. 上海, 上海科學技術出版社. p 337, 1986
  21. 동식, 김근하, 김영호, 최승문. 고려치료경험 내과편. 의학과학출판사. p 71, 76, 114, 122, 132, 166, 345, 386, 1996
  22. 최진규. 약이 되는 우리풀.꽃.나무2. 서울, 한문화. pp 141-147, 2003
  23. 고경식. 가을에 꽃 피는 야생식물. 서울, 일진사. p 56, 2004
  24. 李時珍. 本草綱目. 北京, 人民衛生出版社. p 1366, 1982
  25. 李梴. 編注 醫學入門. 서울, 醫聖堂. p 331, 1994
  26. 寇宗奭. 本草衍義. 서울, 醫聖堂. p 74, 1994
  27. Su-Noh Ryu, Sun-Zik Park, Hong-Yeol Kim. Antioxidant Activity in Rice Cultivar, Wild Rice, and Barley. Korean J Crop Sci 47(1):54-61, 2002
  28. 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
  29. Levine, A.J. The p53 tumor suppressor gene and product. Cancer Surveys. 12, 59-79, 1992
  30. Miyashita, T., Harigai, M., Hanada, M., Reed, J.C. Identification of a p53-dependent negative response element in the Bcl-2 gene. Cancer Res 54, 3131-3135, 1994
  31. El-deiry, W.S., Harper, J.W., O'Connor, P.M., Velculescu, V.E., Canman, C.E., Jackman, J., Pietenpol, J.A., Brrrell, M., Hill, D.E., Wang, Y, Wiman, K.G., Mercer, W.E., Kastan, M.B., Kohn, K.W., Elledge, S.J., Kinzler, K.W., Vogelstein, B. Waf1/CIP1 is induced in p53-mediated G1 arrest and apoptosis. Cancer Res 54, 1169-1174, 1994
  32. Harper, J.W., Adami, G.R., Wei, N., Keyomarsi, K., Elledge, S.J. The p21 cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell 75, 805-816, 1993 https://doi.org/10.1016/0092-8674(93)90499-G
  33. El-Deiry, W.S., Tokino, T., Velculescu, V.E., Levy, D.B., Parsons, R., Trent, J.M., Lin, D., Mercer, W.E., Kinzler, K.W., Vogelstein, B. Waf1, a potential mediator of p53 tumor suppression. Cell 75, 817-825, 1993 https://doi.org/10.1016/0092-8674(93)90500-P
  34. Nakanishi, M., Adami, G.R., Robetorye, R.S., Noda, A., Venable, S.F., Dimitrov, D., Pereira-Smith, O.M., Smith, J.R. Exit from G0 and entry into the cell cycle of cells expressing p21Sdi1 antisense RNA. Proc Natl Acad Sci USA 92, 4352-4356, 1995
  35. Itoh, N, Tsujimoto, Y., Nagata, S. Effect of bcl-2 on Fas antigen-mediated cell death. the journal of immunology 151(2):621-627, 1993
  36. Schlesinger, P.H., Gross, A., Yin, A.M., Yamanoto, K,. Saito, M., Waksman, G,. Korsmeyer, S.J. Comparison of the ion channol characteristics of proapoptotic BAX and antiapoptotic BCL-2. Proc Natl Acad Sci USA 94(21):11357-11362, 1997
  37. Tu, Y,. Feng-hao Xu, Jin Liu, Versco, R., Berenson, J., Fady, C., Lichtanstein, A. Upregulated Expression of Bcl-2 in multiple myeloma cells induced by exposure to Doxorubicin, Etoposide, and Hydrogen peroxide. Blood 88(5):1805-1812, 1996
  38. Lisovsky, M., Estrov, Z., Zhang, X., Consoli, U., Sanchez-Williams, G., Snell, V., Munker, R., Goodacre, A., Savchenko, V., Andreeff, M. Flt3 ligand stimulates proliferation and inhibits apoptosis of acute myeloid leukemia cells : Regulation of Bcl-2 and Bax. Blood 88(10):3987-3997, 1996
  39. Findly. H/W., Gu. L., Yeager. A.M., Zho, M. Expressio and regulation of Bcl-2, Bcl-xL, Bax correlate with p53 status and sensitivity to apoptosis in childhood acute lymphoblastic leukemia. Blood 89(8):2986-2993, 1997