아밀로이드 베타에 의한 신경세포 사멸과 이에 대한열충격 단백질 90의 세포 보호효과
Protective Effect of HSP90 on Neuronal Cell Death-inducedby β-Amyloid Peptide
0
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
본 연구에서는 열충격 단백질 90이 아밀로이드 베타에 의한 신경세포 사멸에 대해 나타내는 보호효과의 분자적 기전을 알아보고자 하였다. 사람의 신경모세포종 SK-N-SH 세포를 이용하였고, 실험을 위하여 AlamarBlue assay, Western blotting assay, Immunofluorescence assay 등을 수행하였다. 그 결과 아밀로이드 베타를 세포에 처리한 경우 세포 사멸이 일어났으며, 이 때 온화한 열충격에 의해 HSP90 의 발현이 증가할 경우 신경세포 사멸이 저해되는 것을 확인하였다. 이 때 존재하는 HSP90의 기능이 무엇인지 알아보기 위해 HSP90의 세포 내 위치를 확인하였는데, 온화한 열충격이 주어질 때 HSP90은 사립체 내에서 특히 발현이 증가하는 것을 관찰할 수 있었다. 또한 HSP90의 발현이 증가하거나 외부에서 처리해 주는 경우 Bcl-2의 발현이 증가하였고, 반대로 HSP90 antisense oligonucleotide를 처리하여 HSP90의 발현을 억제시킨 경우 Bcl-2의 발현이 감소하였다. 그러나 Bcl-2와는 다르게 Bax의 경우는 HSP90의 발현 증감과는 상관이 없었다. 더욱이 HSP90의 발현이 증가한 경우 아밀로이드 베타에 의해 감소되는 사립체 막전위차가 대조군과 유사한 수준으로 유지되는 것을 관찰할 수 있었고, 반대로 HSP90 antisense oligonucleotide를 처리한 경우 아밀로이드 베타에 의한 사립체 막전위차 감소를 더욱 크게 증가시키는 것을 확인할 수 있었다. 이러한 결과는 HSP90이 chaperone 단백질로서 신경독성물질에 의해 나타나는 신경세포 손상을 효과적으로 저 해하고, 세포를 보호하고 있음을 보여준다.
Abstract
In the present study, we determined the protective mechanism of HSP90 against neuronal cell death induced by Aβ. For the evaluation of protective role of HSP90, we used human neuroblastoma SK-N-SH cell lines, examined AlamarBlue assay, Western blot analysis and immunofluorescence assay. Incubation of SK-N-SH cells with Aβ significantly induced neuronal cell death. However, HSP90 induced by mild heat shock could attenuate neuronal apoptosis in Aβ treated condition. To identify the role of HSP90, we determined localization of HSP90 in SK-N-SH cells. Interestingly, HSP90 was increased and localized in mitochondria as treatment of mild heat shock. Also, treatment or increase of HSP90 largely elevated level of Bcl-2 expression, whereas inhibition of HSP90 with HSP90 antisense oligonucleotide significantly decreased Bcl-2 expression. In contrast to Bcl-2, Bax expression was regulated independently by HSP90. Moreover, increase of HSP90 could attenuate collapse of mitochondrial membrane potential induced by Aβ. However, HSP90 antisense oligonucleotide largely increase breakdown of mitochondrial membrane potential induced by Aβ. These data suggest that HSP90 as chaperone protein significantly attenuates neuronal damage and protects neuroanl cells from neurotoxin such as Aβ.
참고문헌 (34)
- Basso AD, "Akt forms an intracellular complex with heat shock protein 90 (Hsp90) and Cdc37 and is destabilized by inhibitors of Hsp90 function", J Biol Chem, 277, pp.39858~39866 (2002)
- Beere HM, "Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome", Nat Cell Biol, 2, pp.469~475 (2000)
- Behl C, "Hydrogen peroxide mediates amyloid beta protein toxicity", Cell, 77, pp.817~827 (1994)
- Forloni G, "Amyloid in Alzheimer’s disease and prion-related encephalopathies: studies with synthetic peptides", Prog Neurobiol, 49, pp.287~315 (1996)
- Frautschy SA, "Effects of injected Alzheimer beta-amyloid cores in rat brain", Proc Natl Acad Sci U S A, pp.8362~8366 (1991)
- Geldmacher DS, "Evaluation of dementia", N Engl J Med, 335, pp.330~336 (1996)
- Gotoh T, "hsp70-DnaJ chaperone pair prevents nitric oxide- and CHOP-induced apoptosis by inhibiting translocation of Bax to mitochondria", Cell Death Differ, 11, pp.390~402 (2004)
- Hofman A, "Atherosclerosis, apolipoprotein E, and prevalence of dementia and Alzheimer’s disease in the Rotterdam Study", Lancet, 349, pp.151~154 (1997)
- Kang J, "The precursor of Alzheimer’s disease amyloid A4 protein resembles a cell-surface receptor", Nature, 325, pp.733~736 (1987)
- Kowall NW, "An in vivo model for the neurodegenerative effects of beta amyloid and protection by substance P", Proc Natl Acad Sci U S A, pp.7247~7251 (1991)
- Lee MW, "Down-regulated reactive oxygen species by HSP90 in 3HK-induced SKN-SH cell death", J Thermal Biol, 30, pp.43~49 (2005)
- Lee MW, "The protective role of HSP90 against 3-hydroxykynurenine-induced neuronal apoptosis", Biochem Biophys Res Commun, 284, pp.261~267 (2001)
- Lewis J, "Disruption of hsp90 function results in degradation of the death domain kinase, receptor-interacting protein (RIP), and blockage of tumor necrosis factor-induced nuclear factor-kappaB activation.", J Biol Chem, 275, pp.10519~10526 (2000)
- Mattson MP, "Pathways towards and away from Alzheimer’s disease", Nature, 430, pp.631~639 (2004)
- Mattson MP, "beta-Amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to excitotoxicity", J Neurosci, 12, pp.376~389 (1992)
- Mayeux R, "Treatment of Alzheimer’s disease", N Engl J Med, 341, pp.1670~1679 (1999)
- McLellan ME, "In vivo imaging of reactive oxygen species specifically associated with thioflavine S-positive amyloid plaques by multiphoton microscopy", J Neurosci, 23, pp.2212~2217 (2003)
- Pandey P, "Negative regulation of cytochrome c-mediated oligomerization of Apaf-1 and activation of procaspase-9 by heat shock protein 90", EMBO J, 19, pp.4310~4322 (2000)
- Pike CJ, "Neurodegeneration induced by beta-amyloid peptides in vitro: the role of peptide assembly state", J Neurosci, 13, pp.1676~1687 (1993)
- Pratt WB, "The hsp90-based chaperone system: involvement in signal transduction from a variety of hormone and growth factor receptors", Proc Soc Exp Biol Med, 통권 : 217, pp.420~434 (1998)
- Reddy PH, "Are mitochondria critical in the pathogenesis of Alzheimer’s disease?", Brain Res Brain Res Rev, 49, pp.618~632 (2005)
- Reddy PH, "Mapping cellular transcriptosomes in autopsied Alzheimer’s disease subjects and relevant animal models", Neurobiol Aging, 27, pp.1060~1077 (2006)
- Saleh A, "Negative regulation of the Apaf-1 apoptosome by Hsp70", Nat Cell Biol, 2, pp.476~483 (2000)
- Sato S, "Modulation of Akt kinase activity by binding to Hsp90", Proc Natl Acad Sci U S A, 통권 : 97, pp.10832~10837 (2000)
- Selkoe DJ, "Alzheimer’s disease: genes, proteins, and therapy", Physiol Rev, 81, pp.741~766 (2001)
- Shearman MS, "Inhibition of PC12 cell redox activity is a specific, early indicator of the mechanism of beta-amyloid-mediated cell death", Proc Natl Acad Sci U S A, 통권 : 91, pp.1470~1474 (1994)
- Shimoke K, "Nerve growth factor prevents 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced cell death via the Akt pathway by suppressing caspase-3-like activity using PC12 cells: relevance to therapeutical application for Parkinson’s disease", J Neurosci Res, 63, pp.402~409 (2001)
- Sisodia SS, "Role of the beta-amyloid protein in Alzheimer’s disease", FASEB J, 9, pp.366~370 (1995)
- Soto C, "Structural determinants of the Alzheimer’s amyloid beta-peptide", J Neurochem, 63, pp.1191~1198 (1994)
- Stankiewicz AR, "Hsp70 inhibits heat-induced apoptosis upstream of mitochondria by preventing Bax translocation", J Biol Chem, 280, pp.38729~38739 (2005)
- Takashima A, "Imahori K : Tau protein kinase I is essential for amyloid beta-proteininduced neurotoxicity", Proc Natl Acad Sci U S A, 통권 : 90, pp.7789~7793 (1993)
- Tanzi RE, "Twenty years of the Alzheimer’s disease amyloid hypothesis: a genetic perspective", Cell, 120, pp.545~555 (2005)
- Vassar R, "Beta-secretase cleavage of Alzheimer’s amyloid precursor protein by the transmembrane aspartic protease BACE", Science, 286, pp.735~741 (1999)
- Yankner BA, "Neurotrophic and neurotoxic effects of amyloid beta protein: reversal by tachykinin neuropeptides", Science, 250, pp.279~282 (1990)