Protective Effect of Puerariae radix Against Ethanol-induced Apoptosis on Human Neuroblastoma Cell Line SK-N-MC

  • Koo Gyo Sung (Oriental Medicine Research Institute, Kyungwon University) ;
  • Cho Son Hae (Oriental Medicine Research Institute, Kyungwon University) ;
  • Jang Mi Hyean (Department of Physiology. College of Medicine, Kyunghee University) ;
  • Kim Chang Ju (Department of Physiology. College of Medicine, Kyunghee University) ;
  • Kim Ee Hwa (Department of Meridinology, College of Oriental. Medicine, Semyung University) ;
  • Lee Choong Yeol (Oriental Medicine Research Institute, Kyungwon University)
  • Published : 2002.06.01

Abstract

To investigate whether Puerariae radix (PR) possesses protective effect against ethanol (EtOH)-inducecl apoptosis in the central nervous system, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, flow cytometric analysis, DNA fragmentation assay. and reverse transcription-polymerase chain reaction (RT-PCR) were performed on human neuroblastoma cell line SK-N-MC. Morphological and biochemical analyses demonstrated that SK-N-MC cells treated with EtOH exhibit classical apoptotic features. On the other hand, cells pre-treated with PR prior to EtOH exposure showed decreased occurrence of classical apoptotic features. In addition, it was shown that PR pre-treatment inhibits EtOH-induced increases in the levels of mRNA expression of bax and caspase-3, while it further enhances the level of bcl-2 expression. These results suggest that PR may exert protective effects against EtOH-induced apoptosis in human neuroblastoma cells.

Keywords

References

  1. Proc Natl Acad Sci USA v.93 Potentiation of the bioavailability of daidzin by an extract of Radix puerariae Keung, W.M.;Lazo, O.;Kunze, L.;Vallee, B.L.
  2. Proc Natl Acad Sci USA v.90 Daidzin: A potent, selective inhibitor of human mitochondrial aldehyde dehydrogenase Keung, W.M.;Vallee, B.L.
  3. J Chromatogr A v.802 Analysis of Puerariae radixand its medicinal preparations by capillary electrophoresis Wang, C.Y.;Huang, H.Y.;Kuo, K.L.;Hsieh,Y.Z.
  4. Proc Natl Acad Sci USA v.95 Daidzin and its antidipsotropic analogs inhibit serotonin anddopamine metabolism in isolated mitochondria Keung, W.M.;Vallee, B.L.
  5. Proc. Natl Acad Sci USA v.94 Daidzin inhibits mitochondrial aldehyde dehydrogenase and suppresses ethanol intake of Syrian golden hamsters Keung, W.M.;Klyosov, A.A.;Vallee, B.L.
  6. Teratology v.40 Differential neuronal loss following early postnatal alcohol exposure Pierce, D.R.;Goodlett, C.R.;West, J.R.
  7. Neurotoxicol Teratol v.19 Ethanol-induced cell death in cultured rat astroglia Holownia, A.;Ledig, M.;Mnez, J.F.
  8. Dev Brain Res v.119 Glial-derived neurotrophic factor (GDNF) prevents ethanol-induced apoptosis and JUN kinase phosphorylation McAlhany Jr, R.E.;West, J.R.;Miranda, R.C.
  9. J Hepatol v.20 Chronic ethanol feeding increases apoptosis and cell proliferation in rat liver Baroni, G. S.;Marucci, L.;Benedetti, A.;Mancini, R.;Jezequel, A.M.;Orlandi, F.
  10. Alcohol Clin Exp Res v.17 Ethanol increases apoptotic cell death of thymocytes in vitro Ewald, S.J.;Shao, H.
  11. Science v.287 Ethanol-induced apoptotic neurodegeneration and fetal alcohol syndrome Ikonomidou, C.;Bittigau, P.;Ishimaru, M.J.;Wozniak, D.F.;Koch, C.;Genz, K.;Price, M.T.;Strfovska, V.;Horster, F.;Tenkova, T.;Dikranian, K.;Olney, J.W.
  12. Cancer Research v.59 no.Supplement Bcl-2gene family and the regulation of programmed cell death Korsmeyer, S.J.
  13. J Neurosci Res v.48 Activation of CPP32 during apoptosis of neurons and astrocytes Keane, R.W.;Srinivasan, A.;Foster, L.M.;Testa, M.P.;rd, T.;Nonner, D.;Wang, H.G.; Reed, J.C.;Bredesen, D.;Kayalar, B.C.
  14. Mol Brain Res v.48 Aberrant expression of bcl-2 gene family in Down's syndrome brains Sawa, A.;Oyama, F.;Cairns, N.J.;Amano, N.;Matsushita, M.
  15. Cell v.74 Bcl-2 heterodimerizes in vivo with a conserved homolog, bax, that accelerates programmed cell death Oltvai, Z.N.;Milliman, C.L.;Korsmeyer, S.J.
  16. J Neurosci v.18 Staurosporine-induced apoptosis of cultured rat hippocampal neurons involves caspase-1-like proteases as upstream initiators and increased production of superoxide as a main downstream effector Krohn, A.J.;Preis, E.;Prehn, J.H.M.
  17. Biochem J v.326 Caspases: the executioners of apoptosis Cohen, G.M.
  18. Jpn J Pharm v.87 Protective effects of Puerariae flos against ethanol-induced apoptosis on human neuroblastoma cell line SK-N-MC Jang, M. H.;Shin, M. C.;Kim, Y. J.;Chung, J. H.;Yim, S. V.;Kim, E. H.;Kim, Y.; Kim, C.J.
  19. Leuk Res v.24 Indomethacin induces poptosis and inhibits proliferation in chronic myeloid leukemia cells Zhang, G.S.;Tu, C.Q.;Zhang, G.Y.;Zhou, G.B.;Zheng, W.L.
  20. Exp Hematol v.28 Expression and activation of caspase-3/CPP32 in CD34+ cord blood cells is linked to apotosis after growth factor withdrawal Wang, L.S.;Liu, H.J.;Xia, Z.B.;Broxmeyer, H.E.;Lu, Li.
  21. Cancer Invest v.10 The origins of DNA breaks: a consequence of DNA damage, DNA repair or apoptosis? Eastman, A.;Barry, M.A.
  22. Eur J Pharm v.385 Enhanced caspase activity during ethanol-induced apoptosis in rat cerebellar granule cells Oberdoerster, J.;Rabin, R.A.
  23. J Neurosci v.19 Bax-dependent caspase-3 activation is a key determinant in p53-induced apoptosis in neurons Cregan, S.P.;MacLaurin, J.G.;Craig, C.G.;Robertson, G.S.;Nicholson, D.W.;Park, D.S.;Slack, R.S.
  24. Nature v.387 Double identity for proteins of the bcl-2 family Reed, J.C.
  25. Science v.258 Prevention of programmed cell death of sympathetic neurons by the bcl-2 proto-oncogene Garcia, I.;Martinou, I.;Tsujimoto, Y.;Martinou, J. C.
  26. Brain Res v.817 Bcl-2 overexpression protects the neonatal cerebellum from ethanol neurotoxicity Heaton, M.B.;Moore, D.B.;Paiva, M.;Gibbs, T.;Bernard, O.
  27. Met Brain Dis v.9 Fetal alcohol syndrome: the vulnerability of the developing brain and possible mechanisms of damage West, J. R.;Chen, W. J.;Pantazis, N. J.
  28. J Cell Biol v.139 Bax deletion further orders the cell death pathway in cerebellar granule cells and suggests a caspase-independent pathway to cell death Miller, T.M.;Moulder, K.L.;Knudson, C.M.;Creedon, D.J.;Deshmukh, M.; Korsmeyer, S.J.;Johnson Jr, E.M.
  29. Gene Dev v.12 Essential contribution of caspase-3/CPP32 to apoptosis and its associated nuclear changes Woo, M.;Hakem, R.;Soengas, M. S.;Duncan, G. S.;Shahinian, A.;Kgi, D.;Hakem, A.;McCurrach, M.;Khoo, W.;Kaufman, S.A.;Senaldi, G.;Howard, T.;Lowe, S.W.;Mak, T.W.
  30. Alcohol Clin Exp Res v.23 Modulation of caspase-3 activity and Fas ligand mRNA expression in rat liver cells in vivo by alcohol and lipopolysccharide Deaciuc, I.V.;Fortunato, F.;D'Souza, N.B.;Hill, D.B.;Schmidt, J.;Lee, E.Y.;McClain, C.J.