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Epigallocatechine gallate에 의한 amyloid β peptide 유도성 신경세포독성의 억제

Inhibition of Amyloid β Peptide-induced Neuronal Cytotoxicity by EGCG

이은주|김현진|김은철|정지연|김원재|김민석|김선헌
대한체질인류학회|2005.06|해부·생물인류학 (Anatomy & Biological Anthropology)|제18권 제2호|pp. 139-147|DOI 10.11637/aba.2005.18.2.139

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Cytotoxicity세포독성amyloid β peptidePC12 세포epigallocatechine gallateAmyloid β peptidePC12 cellsEpigallocatechine gallate
주제분류해부학

초록

본 연구는 PC12 세포에서 amyloid β peptide (Aβ)에 의한 신경독성의 신호전달과정을 구명하며, 녹차의 주요성분으로 강력한 항산화 효과를 나타내는 epigallocatechin gallate (EGCG)에 의한 신경손상 억제효과를 구명 하기 위해 시행되었다. 흰쥐 갈색세포종에서 유래한 세포주인 PC12 세포를 사용하였으며, MTT 환원능을 이용하여 Aβ의 세포독성을 조사하였고, Diff-Quick 염색법을 이용하여 형태적 변화를 관찰하였다. Aβ에 의한 PC12 세포의 죽음과 이를 억제 하는 EGCG의 작용기전도 신호전달과정을 알기 위해 활성산소종(ROS) 측정, western blot, RT-PCR 등의 방법을 사용하였다. Aβ에 의한 세포독성은 농도 의존적으로 나타났으며 이는 배양 세포의 핵분절화를 비롯한 세포자멸사와 유사한 형태적 변화에 의해 확인되었다. Aβ에 의한 세포독성은 EGCG의 전처치에 의해 효과적으로 억제되었다. 또한 EGCG는 Aβ투여 후 발생하는 활성산소종을 ROS 제거제인 NAC (N-acetyl cystein)의 처치 수준으로 감소시켰다. 세포죽음의 신호전달과정 중 사립체 의존 경로에 관련된 단백질인 세포질 내 cytochrome c의 발현양은 Aβ 투여후 12시간부터 증가하였으며, EGCG에 의해 현저히 감소하였다. 세포질 내 cytochrome c의 유리를 조절하는 Bax/Bcl-2의 발현양상을 RT-PCR을 통해 확인하였다. Aβ는 Bax의 발현을 증가시켰으나, Bcl-2의 발현에는 영향을 주지 않았으며, EGCG는 Aβ에 의한 Bax의 증가를 억제하였다.이상의 결과로 Aβ에 의한 신경세포 독성은 ROS 및 세포죽음의 사립체 의존 경로에 의한 것으로 보이며, EGCG는 항산화효과 외에도 세포죽음의 사립체 의존 경로와 관련한 단백의 발현 억제를 통해 효과적으로 신경세 포를 보호하는 것으로 생각된다.

Abstract

This study is aimed to investigate the signal transduction pathway of amyloid β peptide (Aβ)-induced neuronal toxicity and the inhibitory effects of epigallocatechin gallate (EGCG), one of the major constituents of green-tea and the potent anti-oxidant, on the nerve cell damage in PC12 cells. Cellular toxicity was estimated by MTT assay and observation of morphological changes in PC12 cells. By using the methods such as measurement of Reactive Oxygen Species (ROS), western blot and RT-PCR, the underlying mechanisms and signal transduction pathway of Aβ-induced neurotoxicity and the inhibitory effects of EGCG were examined. Aβ-induced cellular toxicity was found in a dose dependent manner. This is confirmed by morphological observations of cultured cells such as findings of cell death similar to apoptosis. Aβ-induced neurotoxicity was effectively inhibited by EGCG pretreatment. Moreover, EGCG reduced ROS as same potent as the NAC (N-acetyl cystein), the ROS scavenger. Among the several process of signal transduction for cell death, a intracytoplasmic cytochrome c, the protein associated with the mitochondria-dependent pathway, was increased from 12 hours after Aβ treatment and the increased cytochrome c by Aβ was blocked by EGCG. Expression levels of Bax/Bcl-2 in relation to intracytoplasmic release of cytochrome c were examined by RT-PCR. Aβ up-regulated Bax expression but did not affect Bcl-2 expression. EGCG was found to block the effect of Aβ-induced Bax increase. From these results, it is speculated that Aβ-induced neuronal toxicity may be assumed to be affected by ROS and the mitochondria-dependent pathway of cell death as well. EGCG, besides having the role of anti-oxidant, is found to have a protective effect against Aβ-induced neurotoxicity through the inhibition of the expression of the protein associated with the mitochondria-dependent cell death pathway.

참고문헌 (34)

  1. "Key factors in Alzheimer’s disease", pp.1~11 (2001)
  2. "Mechanism of amyloid beta protein inducedneuronal cell death:current concepts and futureperspectives", 49, pp.125~134 (1997)
  3. "Acetaminophenprotects hippocampal neurons and PC12 culturesfrom amyloid beta-peptides induced oxidative stress andreduces NF-kappaB activation", 41, pp.43~54 (2002)
  4. "Antioxidant capacity of flavanols andgallate esters:pulse radiolysis studies", pp.1413~1426 (1999)
  5. "Mitochondrial redox signaling during apoptosis", 31, pp.327~334 (1999)
  6. "Inhibition of induciblenitric oxide synthase gene expression and enzymeactivity by epigallocatechin gallate", pp.1281~1286 (1997)
  7. "Phosphatidylinositol-3-kinase-Akt kinase and p42", p44 mitogen-activatedprotein kinases mediate neurotrophic and excitoprotectiveactions of a secreted form of amyloid precursor protein, 2002, pp.175, 407~414
  8. "Molecularbasis of Alzheimer’s disease", 57, pp.705~715 (2000)
  9. "and neurotoxicity of the Alzheimer’sAbeta1-42 amyloid peptide and its isoaspartylisomers", pp.953~956 (1999)
  10. "Long-term changes in the aggregation stateand toxic effects of beta-amyloid injected into the ratbrain", 87, pp.349~357 (1998)
  11. "prevents the initial decrease in mitochondrial membranepotential and subsequent reactive oxygen species productionduring tumor necrosis factor alpha-induced apoptosis", pp.5680~5689 (2000)
  12. "Studies on protectivemechanisms of four components of green tea polyphenolsagainst lipid peroxidation in synaptosomes", 1304, pp.210~222 (1996)
  13. "ESR study onthe structure-antioxidant activity relationship of tea catechinsand their epimers", 1427, pp.13~23 (1999)
  14. "The correlation betweenactive oxygens scavenging and antioxidative effects of flavonoids", 16, pp.845~850 (1994)
  15. "Mechanisms of beta-amyloid neurotoxicity perspectives of pharmacotherapy", pp.329~382 (2000)
  16. "Reactive oxygen species as causal agents inthe neurotoxicity of the Alzheimer’s disease-associatedamyloid beta peptide", akse(v m), pp.120~1341996
  17. "Inhibition of mitochondrialrespiration and cyanide-stimulated generation ofreactive oxygen species by selected flavonoids", 47, pp.573~580 (1994)
  18. "Estrogen protects neuronalcells from amyloid beta-induced apoptotic cell death", 12, pp.1965~1970 (2001)
  19. "Interactions of flavonoidsand other phytochemicals with adenosine receptors", 39, pp.781~788 (1996)
  20. "Role of flavonoidsand iron chelation in antioxidant action", pp.437~443 (1994)
  21. "adysfunction of the amyloid precursor protein", pp.54~66 (2000)
  22. "Distinct effects of teacatechins on 6-hydroxydopamine-induced apoptosis inPC12 cells", 397, pp.84~90 (2002)
  23. "Mechanisms of growth inhibition of human lungcancer cell line", pp.639~9 643 (1997)
  24. "Evidence of oxidative stress and in vivoneurotoxicity of beta-amyloid in a transgenic mouse modelof Alzheimer’s disease a chronic oxidative paradigmfor testing antioxidant therapies in vivo", pp.871~877 (1998)
  25. "Neuroprotective effect ofgarlic compounds in amyloid-beta peptide-induced apoptosisin vitro", 8, pp.~337 (2002)
  26. "Flavonoids as antioxidant agents importance of theirinteraction with biomembranes", pp.481~486 (1995)
  27. "Electrophysiologicalstudy of a novel large pore formed by bax and thevoltage-dependent anion channel that is permeable tocytochrome c", 275, pp.12321~12325 (2000)
  28. "Cytochrome c release into cytosol with subsequentcaspase activation during warm ischemia in ratliver", 281 (2001)
  29. "Regulation of hydrogenperoxide production by brain mitochondria by calcium andBax", 83, pp.220~228 (2002)
  30. "beta-Amyloid-induced neuronalapoptosis requires c-Jun N-terminal kinase activation", 77, pp.157~164 (2001)
  31. "The Ginkgo biloba extractEGb 761 rescues the PC12 neuronal cells from beta-amyloid-induced cell death by inhibiting the formation of beta-amyloid-derived diffusible neurotoxic ligands", pp.181~190 (2001)
  32. "Involvement of apoptosisinducingfactor in neuronal death after hypoxia-ischemiain the neonatal rat brain", pp.306~317 (2003)
  33. "cytochrome cmultimeric complex is a functional apoptosome that activatesprocaspase-9", pp.11549~11556 (1999)
  34. "Involvementof caspase-3 and p38 mitogen-activated proteinkinase in cobalt chloride-induced apoptosis in PC12cells", 67, pp.837~843 (2002)

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아밀로이드베타 펩타이드 유도성 신경세포독성에 대한 녹차 추출물의 보호 효과김영인, 김재겸, 박정윤, 이승철, 정창호, 조성환, 최성길, 최수정, 허호진한국식품저장유통학회 · Food Science and Preservation2008 · 제15권 제5호PC12세포에서 CoCl2 유발 세포자멸사에 대한epigallocatechin-gallate의 역할김선미, 김원재, 모현철, 양규호, 최남기대한소아치과학회 · 대한소아치과학회지2006 · 제33권 제1호PC12 세포에서 산화스트레스에 의한 베타-아밀로이드 펩티드의 세포 독성김병학, 김성용, 김재룡, 김정희, 김찬석, 김태정, 이재규, 하정상대한신경과학회 · 대한신경과학회지2003 · 제21권 제4호Green tea polyphenol (-)-epigallocatechin 3-gallate prevents ultraviolet--induced apoptosis in PC12 cells김영, 강인철, 김옥준, 김원재, 김윤정, 박삼영, 우수미, 정지연, Bangrong Cai대한구강생물학회 · International Journal of Oral Biology2020 · 제45권 제4호일시적 국소허혈에 의해 유발된 흰쥐 대뇌 신경세포 손상에 대한 EGCG (epigallocatechin gallate)의신경보호효과김범생, 박정욱, 이광수, 최영빈, 한시령대한신경과학회 · 대한신경과학회지2003 · 제21권 제4호Epigallocatechin gallate attenuates L-DOPA-induced apoptosis in rat PC12 cells이명구, 이명렬, 이재준, 최은주한국영양학회 · Nutrition Research and Practice2013 · 제7권 제4호

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