The Effect of Woohwangcheongsim-won for Delayed Neuronal Death in OGD(Oxygen-Glucose Deprivation) Model

배양 대뇌신경세포의 저당-저산소증 모델에서 우황청심원에 의한 세포사 방지 연구

  • 원철환 (동국대학교 한의과대학 내과학교실) ;
  • 정승현 (동국대학교 한의과대학 내과학교실) ;
  • 신길조 (동국대학교 한의과대학 내과학교실) ;
  • 문일수 (동국대학교 의과대학 해부학교실) ;
  • 이원철 (동국대학교 한의과대학 내과학교실)
  • Published : 2002.12.01

Abstract

Objectives: The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won and to study the mechanism for neuronal death protection in OGD (oxygen-glucose deprivation) model with embryonic day 20 (E20) cortical cells of a rat (Sprague Dawley). Methods: E20 cortical cells were dissociated in neurobasal media and grown for 14 days in vitro (DIV). On 14 DIV, Woohwangcheongsim-won was added to the culture media for 72 hrs. On 17 DIV, cells were given an oxygen-glucose deprivation shock (2hrs and 4hrs) and further incubated in normoxia for another three days. On 20 DIV, Woohwangcheongsim-won's effects for neuronal death protection were evaluated by LDH assay and the mechanisms were studied by Bcl-2, Bak, Bax, caspase family. Results & Conclusions: 1. This study indicates that Woohwangcheongsim-won's effects for neuronal death protection in OGD model is confirmed by LDH assay in culture method of embryonic day 20(E20) cortical neuroblasts. 2. Woohwangcheongsim-won's mechanisms for neuronal death protection in OGD model are to restrain inflow of cytochrome c into cellularity caused by Bcl-2 increase (2hrs and 4hrs), to reduce the caspase cascade initiator caspase-8 (4hrs).

Keywords

References

  1. Science v.267 Apoptosis in the pathogenesis and treatment of disease Thompson CB
  2. Dev. Biol. v.138 Naturally occuring and induced neuronal cell death in the chick embryo in vivo requires protein and RNA synthesis. evidence for the role of ccell death genes. Oppenheim RV;Prevette D;Tytell M;S Homma
  3. Gene. Devel. v.12 Proteases to die for Cryns V;J Yuan
  4. Annu. Rev. Physiol. v.60 The mitochondrial death/life regulator in apoptosis and necrosis Kroemer G;Dallaporta B;M Resche-Rigon
  5. 중풍학술대회논문집 v.1 중풍(뇌졸중) 김영석
  6. 東醫寶鑑 許浚
  7. 濟衆新編 康命吉
  8. 대한한의학회지 v.15 no.1 의식장애를 동반한 졸중풍 급성기 환자에서 우황청심원 투여에 관한 고찰 김동웅;오승환;안일회;이언정;이형균;이승무 등
  9. J. Neurosci. Res v.35 Optimized survival of hippocampal neurons in B27-supplemented Neurobasal, a new serum-free medium combination Brewer GJ;Torricelli JR;Evege EK;Price PJ
  10. 우황청심원이 저산소증으로 유발된 지연성 신경세포사에 미치는 영향 김민석
  11. Neuron v.1 Glutamate Neurotoxity and Diseases of the Nervous System Dennis W. Choi.
  12. Brain Res. v.239 no.1 Delayed neuronal death in the gerbil hippocampus following ischemia Kirino T
  13. J Neurochem v.68 no.4 Evidence that the early loss of membrane protein kinase C is a necessary step in the excitatory amino acid-induced death of primary cortical neurons Durkin JP;Tremblay R;Chakravarthy B;Mealing G;Morley P;Small D(et el)
  14. Prog Neurobiol v.62 no.3 Mechanisms underlying hypoxia-induced neuronal apoptosis Banasiak KJ;Xia Y;Haddad GG
  15. J Neurochem v.66 no.3 An early loss in membrane protein kinase C activity precedes the excitatory amino acid-induced death of primary cortical neurons Durkin JP;Tremblay R;Buchan A;Blosser J;Chakravarthy B;Mealing G(et el)
  16. 경희의학 v.19 우황청심원과 소합향원의 효능에 대한 연구 남상경;이경섭
  17. 청강의감 이종형
  18. 생약학회지 v.18 no.4 우황청심원이 순환기계에 미치는 영향 홍남두;김남재;김규섭
  19. 약학회지 v.41 no.6 우황청심원의 뇌허혈 및 중추신경계에 대한 약효 조태순;이선미;이은방;조성익;김용기;신대희;박대규
  20. 우황청심원이 인체내피세포의 NOS 및 세포부착인자 유전자 발현에 미치는 영향에 관한 분자생물학적 연구 문상관
  21. 우황청심원이 eNOS, 세포유착분자, TNF$-\alpa$ 및 p53 유전자 발현에 미치는 영향 한창호
  22. 대한한의학회지 v.22 no.1 우황청심원이 중대뇌동맥 결찰로 유발된 뇌허혈에 미치는 영향 조규선;정승현;신길조;이원철
  23. 대한한방내과학회지 v.22 no.2 우황청심원이 저산소증 유발 배양신경세포에 미치는 영향 조규선;정승현;신길조;문일수;이원철
  24. Masui v.47 no.6 The pathophysiology of ischemic neuronal injury. an overview Iijima T
  25. J Neurosci v.6 no.11 Effects of ischemia-like conditions on cultured neurons, protection by low Na+, low Ca2+ solutions Goldberg WJ;Kadingo RM;Barrett JN
  26. J Neurochem v.59 no.3 Regulatory interactions among axon terminals affecting the release of different transmitters from rat striatal slices under hypoxic and hypoglycemic conditions Milusheva E;Doda M;Pasztor E;Lajtha A;Sershen H;Vizi ES
  27. Brain Res. v.607 no.1;2 Effect of varying levels of glucose on oxygen deprivation-induced delayed neuronal cell death in primary cerebrocortical cultures Birrell GJ;Marcoux FW
  28. J Neurophysiol v.85 no.3 Hypoglycemia enhances ionotropic but reduces metabotropic glutamate responses in substantia nigra dopaminergic neurons Mercuri NB
  29. Neurosci Lett. v.309 no.3 Lowering glucose concentrations increases cytosolic Ca2+ in orexin neurons of the rat lateral hypothalamus Yada T
  30. Biochem Soc Trans. v.29 no.6 Caspases: cellular demolition experts Creagh EM;Martin SJ
  31. Science v.281 no.5381 Caspases: enemies within Thornberry NA;Lazebnik Y
  32. J Neurosci Res v.64 no.2 TNF-alpha stimulates caspase-3 activation and apoptotic cell death in primary septo-hippocampal cultures Zhao X;Bausano B;Pike BR;Newcomb-Fernandez JK;Wang KK;Shohami E;Ringger NC;DeFord SM;Anderson DK;Hayes RL
  33. Science v.281 no.5381 Mitochondria and apoptosis Green DR;Reed JC
  34. Cell v.86 no.1 Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrsome c Liu X;Kim CN;Yang J;Jemmerson R;Wang X
  35. J Biol Chem v.274 no.25 Cytochrsome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation Saleh A;Srinivasula SM;Acharya S;Fishel R;Alnemri ES
  36. Genes Dev. v.13 no.24 Caspase-9 and APAF-1 form an active holoenzyme Rodriguez J;Lazebnik Y
  37. Genes Dev. v.13 no.15 BCL-2 family members and the mitochondria in apoptosis Gross A;McDonnell JM;Korsmeyer SJ
  38. Genes Dev. v.14 no.16 tBID, a membrane-targeted death ligand, oligomerizes BAK to release cytochrsome c Wei MC;Lindsten T;Mootha VK;Weiler S;Gross A;Ashiya M(et al)
  39. J Cell Biol. v.144 no.5 Bid-induced conformational change of Bax is responsible for mitochondrial cytochrsome c release during apoptosis Desagher S;Osen-Sand A;Nichols A;Eskes R;Montessuit S;Lauper S(et al)
  40. Science v.275 no.5303 The release of cytochrsome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis Kluck RM;Bossy-Wetzel E;Green DR;Newmeyer DD