The influence of endogenous β-amyloid on mitochondria and expression of heat shock protein in Neuro 2a cells bearing the amyloid precursor protein and primary neuron cell

• Main Authors: Sheng-Jie You, 游勝傑
• Other Authors: 楊婷婷
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/01934809303036228767

碩士 === 中國醫藥大學 === 醫學研究所碩士班 === 95 === Aβ accumulation to oligomers is a kind of stress which can cause cell damage by inducing oxidative stress in vitro studies. Heat shock proteins (HSPs) induced by stress. The relationship between endogenous Aβ and HSP expression remains uncleaer. In our study, APPwt transfected neuro 2a (N-wt) and APPsw gene transfected neuro 2a (N-sw) cells and primary neuron were used. We evaluated viability of N-wt and N-sw via trypan blue stain. We found the cell viability was decreased compared with control. Result of cell cycle showed that sub G1 peak of N-wt and N-sw was increased compared with control. DNA fragmentations showed apoptotic features in N-wt, N-sw. JC-1 stain(mitochondrial membrane potential), mitotracker stain(ROS level), Fluo-3A (Ca2+ level) stain and cytochrome c oxidase assay showed mitochondrial function impared. Western blot showed expression of pro-apoptotic protein Bid and cytochrome c increased while expression of anti-apoptotic protein Bcl-2 compared with control, respectively. Increased expression of HSP70, HSP60, and HSP27 were in N-wt, N-sw while HSP90, HSP32 were decreased in N-wt, N-sw compared with control, respectively. Similar results were found in primary neuron; Aβs decreased cell viability of neuronal cell and influenced mitochondria function. Increased expression of pro-apoptotic related protein, Bid, cytochrome c while expression of anti-apoptotic protein Bcl-2 was decreased compared with control. Increased expression of HSP90, HSP70, HSP60, HSP27 while HSP32 decreased compared with control. This is the first study of the correlation between Aβs and HSP. Aβs is a potent source of oxidative stress, but expression of HSP induced by Aβs remains unclear. We respect expression of HSP up regulated, but HSP 90 in N-wt, N-sw and HSP 32 are down regulated. HSP induced by stress and could protect cell, but in this study, the viability of neurons were decreased compare with control. The neuron protection of HSP while Aβs exists needs further investigation.