본문 바로가기
KERD
논문KCI 등재

Glutamate로 유도된 C6 glial 세포 자멸사에 대한 淸心蓮子飮의 보호효과

Protective Effects of Chungsimyeonja-eum on Glutamate-induced Apoptosis in C6 Glial Cells

안민섭|신용진|권오상|장원석|신선호|하예진|이선아|고석재
대한한방내과학회 (The Society Of Korean Oriental Internal Medicine)|2010.03|대한한방내과학회지 (The Journal of Korean Oriental Internal Medicine)|제31권 제1호|pp. 54-65

2

조회

0

다운로드

3

인용

0

참고문헌

키워드

ApoptosisGlutamateC6 glial cellChungsimyeonja-eum
주제분류한의학

초록

Objective : The water extract of Chungsimyeonja-eum (CSYJE) has traditionally been used in treatments of heart diseases and brain diseases in Oriental medicine. However, little is known about the mechanism by which CSYJE protects neuronal cells from injury damages. Therefore, in this study we attempted to elucidate the mechanism of the cytoprotective effect of the CSYJE extract on glutamate-induced C6 glial cell death. Methods : Cultured cells were pretreated with CSYJE and exposed to glutamate, cell damage was assessed by using MTT assay and propidium iodide (PI), probe 2',7'-dichlorofluorescein diacetate (DCF-DA) staining. Western blotting was performed using anti-procaspase-3 and anti-PARP, respectively. Result : We determined the elevated cell viability by CSYJE extract on glutamate-induced C6 glial cell death. Glutamate induced DNA fragmentation on C6 glial cells but pre-treatment with CSYJE inhibited DNA fragmentation. One of the main mediators of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pre-treatment with CSYJE inhibited this ROS generation from glutamate-stimulated C6 glial cells. Also, we identified that the ROS-induced DCF-DA green fluorescence was reduced by CSYJE pre-treatment. The critical markers of apoptotic cell death are the cleavages of procaspase-3 protease and PARP proteins, so we checked the expression level and cleavages of procaspase-3 protease and PARP proteins. Glutamate-treated C6 glial cells showed the cleavages of procaspase-3 protease and PARP proteins and followed the reduction of expression of these proteins. Conclusion : These findings indicate that CSYJE may prevent cell death from glutamate-induced C6 glial cell death by inhibiting the ROS generation and procaspase-3 and PARP expression.

Abstract

Objective : The water extract of Chungsimyeonja-eum (CSYJE) has traditionally been used in treatments of heart diseases and brain diseases in Oriental medicine. However, little is known about the mechanism by which CSYJE protects neuronal cells from injury damages. Therefore, in this study we attempted to elucidate the mechanism of the cytoprotective effect of the CSYJE extract on glutamate-induced C6 glial cell death. Methods : Cultured cells were pretreated with CSYJE and exposed to glutamate, cell damage was assessed by using MTT assay and propidium iodide (PI), probe 2',7'-dichlorofluorescein diacetate (DCF-DA) staining. Western blotting was performed using anti-procaspase-3 and anti-PARP, respectively. Result : We determined the elevated cell viability by CSYJE extract on glutamate-induced C6 glial cell death. Glutamate induced DNA fragmentation on C6 glial cells but pre-treatment with CSYJE inhibited DNA fragmentation. One of the main mediators of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pre-treatment with CSYJE inhibited this ROS generation from glutamate-stimulated C6 glial cells. Also, we identified that the ROS-induced DCF-DA green fluorescence was reduced by CSYJE pre-treatment. The critical markers of apoptotic cell death are the cleavages of procaspase-3 protease and PARP proteins, so we checked the expression level and cleavages of procaspase-3 protease and PARP proteins. Glutamate-treated C6 glial cells showed the cleavages of procaspase-3 protease and PARP proteins and followed the reduction of expression of these proteins. Conclusion : These findings indicate that CSYJE may prevent cell death from glutamate-induced C6 glial cell death by inhibiting the ROS generation and procaspase-3 and PARP expression.

참고문헌 (47)

  1. Cunningham LM, "A review of assistive technologies for people with Parkinson's disease", Technol Health Care, 17(3), pp.269~279 (2009)
  2. 나영설, "최근 뇌졸중에 대한 역학적 고찰", pp.280~286 (1991), 경희의학
  3. Kovari E, "Neuropathology of Lewy body disorders", Brain Res Bull., 80(4), pp.203~210 (2009)
  4. Zuccato C, "Brain-derived neurotrophic factor in neurodegenerative diseases", Nat Rev Neurol., 5(6), pp.311~322 (2009)
  5. 대한신경외과학회, "신경외과학", pp.15~ (1997), 중앙문화사
  6. Witte OW, "Functional differentiation of multiple perilesional zones after focal cerebral ischemia", J. Cereb Blood Flow Metab., 20(8), pp.1149~1165 (2000)
  7. Graham SH, "Programmed cell death in cerebral ischemia", J. Cereb Blood Flow Metab, 21(2), pp.99~109 (2001)
  8. Cho Y, "Uptake of glutamate and cystein in C6 glioma cells and in cultured astrocytes", J. Neurochem., 55, pp.2091~2097 (1990)
  9. Landucci E, "Involvement of endocannabinoid signaling in the neuroprotective effects of subtype 1 metabotropic glutamate receptor antagonists in models of cerebral ischemia", Int Rev Neurobiol., 85, pp.337~350 (2009)
  10. Dean OM, "A role for glutathione in the pathophysiology of bipolar disorder and schizophrenia: Animal models and relevance to clinical practice", Curr Med Chem., 16(23), pp.2965~2976 (2009)
  11. Milanese M, "Glutamate release from astrocytic gliosomes under physiological and pathological conditions", Int Rev Neurobiol, 85, pp.295~318 (2009)
  12. Gao Z, "Protective effects of flavonoids in the roots of Scutellaria baicalensis Georgi against hydrogen peroxide-induced oxidative stress in SH-SY5Y cells", Pharmacol Res., 43(2), pp.173~178 (2001)
  13. Halliwell B, "Protection aganist oxidants in biological systems: the superoxide theory of oxygen toxicity. In Free radicals in biology and medicine", pp.86~89 (1999), Clarendon Press
  14. 劉景源點校, "太平惠民和劑局方", pp.196~ (1985), 人民衛生出版社
  15. 황도연, "방약합편", pp.213~214 (2002), 영림사
  16. 윤용갑, "동의방제와 처방해설", pp.790~ (2004), 의성당
  17. 권영철, "중풍치료에 응용되는 淸心蓮子飮에 관한 문헌적 고찰", 대한한의학회지, 6(1), pp.55~59 (1985)
  18. 박순일, "淸心蓮子飮과 星香正氣散이 Streptozotocin誘發 糖尿흰쥐의 腦虛血 損傷에 미치는 影響", 대한한의학회지, 28(3), pp.216~231 (2007), 대한한의학회
  19. 윤정환, "淸心蓮子飮의 구속 스트레스 및 절개 통증에 대한 항스트레스 작용과 진통효과", 박사, pp.2006
  20. Hutchins JB, "Why neurons die: cell death in the nervous system", Anat Rec, 253(3), pp.79~90 (1997)
  21. Fischer U, "New approaches and therapeutics targeting apoptosis in disease", Pharmacol Rev., 57(2), pp.187~215 (2005)
  22. Uversky VN, "Intrinsic disorder in proteins associated with neurodegenerative diseases", Front Biosci., 14, pp.5188~5238 (2009)
  23. Nicholls DG, "Mitochondrial dysfunction and glutamate excitotoxicity studied in primary neuronal cultures", Curr Mol Med., 4, pp.149~177 (2004)
  24. Farooqui T, "Aging: an important factor for the pathogenesis of neurodegenerative diseases", Mech Ageing Dev., 130, pp.203~215 (2009)
  25. Berman SB, "Dopamine oxidation alters mitochondrial respiration and induces permeability transition in brain mitochondria, implications for Parkinson’s disease", J. Neurochem, 73, pp.1127~1137 (1999)
  26. Halestrap AP, "Mitochondria and cell death", Biochem Soc Trans, 28, pp.170~177 (2000)
  27. Tournier C, "Requirement of JNK for stress-induced activation of the cytochrome c-mediated death pathway", Science, 288, pp.870~874 (2000)
  28. Luo Y, "Intrastriatal dopamine injection induces apoptosis through oxidation-involved activation of transcription factors AP-1 and NF-kappaB in rats", Mol Pharmacol., 56, pp.254~264 (1999)
  29. Maalouf M, "Ketones inhibit mitochondrial production of reactive oxygen species production following glutamate excitotoxicity by increasing NADH oxidation", Neuroscience, 145, pp.256~264 (2007)
  30. 신민교, "임상본초학", pp.172~ (2002), 영림사
  31. 이인자, "소갈증 치료제가 실험적 당뇨 흰쥐에 미치는 영향 -청심연자음, 가미조위승기탕, 기국지황탕", 약학회지, 36(4), pp.555~561 (1994)
  32. 김수웅, "淸心蓮子飮이 Streptozocin으로 유발한 백서의 고혈당에 미치는 영향", 동의병리학회지, 6, pp.63~86 (1991)
  33. 이명규, "미세변화 신증후군에서 淸心蓮子飮과 大分淸飮이 면역조절기능에 미치는 영향", 박사, pp.2000
  34. 이상헌, "淸心蓮子飮이 Mouse 유래 C2C12세포주에서 심근세포 손상의 보호효과", 대한한의학회지, 27(3), pp.25~36 (2006), 대한한의학회
  35. 문형철, "人蔘이 중추신경계 손상 동물 모델의 재생에 미치는 영향", 대한침구학회지, 24(6), pp.137~148 (2007), 대한침구학회
  36. 임강현, "새로운 뇌 위축 동물 모델과 그 모델에서의 고려인삼의 보호 효과", 대한본초학회지, 21(4), pp.197~205 (2006), 대한본초학회
  37. 이진우, "연자육의 항우울 효과 및 프로티옴 분석을 통한 기전 연구", 동의생리병리학회지, 20(4), pp.830~843 (2006), 대한동의병리학회
  38. 김효근, "연자육의 6-하이드록시도파민으로 유도된 도파민 세포 독성에 대한 보호효과", 대한본초학회지, 24(2), pp.87~92 (2009), 대한본초학회
  39. 오병열, "加味柴胡湯이 뇌허혈시 Glutamate receptor, free radical 및 뇌손상 보호에 미치는 영향", 대한한의학회지, 25(3), pp.32~44 (2004), 대한한의학회
  40. 강익현, "歸脾湯이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향", 대한한의학회지, 22(4), pp.45~57 (2001)
  41. 정대영, "四物湯이 Glutamate에 의한 해마세포의 손상에 미치는 영향", 대한한의학회지, 30(1), pp.64~75 (2009), 대한한의학회
  42. 정승원, "疎風湯이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향", 동의생리병리학회지, 22(6), pp.1423~1430 (2008), 대한동의병리학회
  43. Bose R, "Ceramide synthase mediates daunorubicin-induced apoptosis: an alternative mechanism for generating death signals", Cell, 82, pp.405~414 (1995)
  44. Brown DG, "Dexamethasone -induced apoptosis involves cleavage of DNA to large fragments prior to internucleosomal fragmentation", J. Biol Chem., 268, pp.3037~3039 (1993)
  45. Budijardjo I, "Biochemical pathways of caspase activation during apoptosis", Annu Rev Cell Dev Biol, 15, pp.269~290 (1999)
  46. Atalay M, "Altered antioxidant enzyme defences in insulin-dependent diabetic men with increased resting and exercise-induced oxidative stress", Acta Physiol Scand, 161(2), pp.195~201 (1997)
  47. Halliwell B, "Role of free radicals and catalytic metal ions in human disease: an overview", Methods Enzymol., 186, pp.1~85 (1990)

관련 논문AI추천

大七氣湯이 Glutamate에 의한 C6 Glial 세포의 Apoptosis에 미치는 영향이인, 고석재, 김혜윤, 방창호, 신선호, 이동엽대한한방내과학회 · 대한한방내과학회지2010 · 제31권 제4호六君子湯이 Glutamate에 의한 C6 신경교세포의 Apoptosis에 미치는 영향고석재, 권영미, 신선호, 신용진, 장원석, 하예진대한한방내과학회 · 대한한방내과학회지2010 · 제31권 제3호疎風湯이 Glutamate에 의한 C6 Glial Cell의 Apoptosis에 미치는 영향김봉상, 문병순, 정승원, 최철원한의병리학회 · 동의생리병리학회지2008 · 제22권 제6호眞武湯이 H_2O_2로 유도된 C6 Glial 세포사에 미치는 영향신선호, 신용진, 유주연, 이숭인, 조문영, 최정훈, 하예진대한한방내과학회 · 대한한방내과학회지2012 · 제33권 제3호Glutamate 로 유도된 C6 glial 세포의 독성에 대한 八物湯의 보호 효과신선호, 신용진한의병리학회 · 동의생리병리학회지2012 · 제26권 제4호H2O2로 유발된 C6 신경교세포 사멸에 대한 총명공진단의 보호 효과신용진, 황규상대한한방내과학회 · 대한한방내과학회지2020 · 제41권 제1호

이 간행물의 다른 논문 (15건)

한국표준질병사인분류중 한방내과영역의 분류체계 개선 및 진단명 구성에 관한 연구이원철pp. 1-10枳椇子 山靑木 配合이 D-galactosamine 유발 백서의 간손상에 미치는 影響문구, 이선아pp. 11-24消平湯이 RIN-m5F 세포에서 인슐린 분비 및 유전자 발현에 미치는 영향조충식, 윤성식pp. 25-39진전 변증에 관한 문헌적 고찰 - 중국 논문 중심으로 -선승호pp. 40-53황기가 흰쥐의 만성 신손상에 미치는 영향이권호, 김용성, 한양희pp. 66-78
원문 보기