本文選題：AD + PS1��； 參考：《華東師范大學》2016年碩士論文

【摘要】：阿爾茲海默癥(Alzheimer disease, AD)是臨床上常見的神經(jīng)退行性疾病之一,認知能力的逐步喪失是其主要的臨床特征,且伴隨大腦中神經(jīng)元胞外神經(jīng)炎斑塊Ap的大量累積和Tau蛋白的過度磷酸化導(dǎo)致的神經(jīng)元纖維纏結(jié),這些病理學變化被認為是大腦神經(jīng)細胞大量凋亡及發(fā)生認知障礙的主要原因。近年來研究顯示神經(jīng)元凋亡還與線粒體和內(nèi)質(zhì)網(wǎng)功能異常有關(guān)。我們前期在PS1/PS2條件性雙敲(PS1/2 conditional double knock out, PS1/2cDKO)小鼠大腦中發(fā)現(xiàn)神經(jīng)元線粒體出現(xiàn)功能和形態(tài)異常；在PS1干擾(即PS功能缺失)的SH-SY5Y細胞模型上,我們則發(fā)現(xiàn)PS1功能缺失誘導(dǎo)的神經(jīng)元凋亡受線粒體PS1-PARL-OPA1信號途徑調(diào)控,即在PS1瞬時干擾細胞模型中過表達PARL后,下降的OPA1便上調(diào)到正常水平,凋亡現(xiàn)象出現(xiàn)減弱。本研究中,我們進一步發(fā)現(xiàn)PS1/2 cDKO小鼠大腦抗凋亡蛋白PARL與其調(diào)控的線粒體融合蛋白OPA1在RNA水平和蛋白水平均表達下調(diào),與PS1干擾的SH-SY5Y細胞模型上觀察到的研究結(jié)果一致。本研究圍繞線粒體PS1-PARL-OPA1凋亡途徑,發(fā)現(xiàn)在PS1瞬時干擾細胞模型中過表達OPA1后并不改變PS1和PARL的低表達,凋亡現(xiàn)象未出現(xiàn)顯著的改變,說明OPA1雖過表達但缺少PARL對其的剪切,仍無法行使其正常生理功能。線粒體和內(nèi)質(zhì)網(wǎng)(Endoplasmic Reticulum, ER)作為兩個獨立的細胞器,雖有不同的分工但在細胞穩(wěn)態(tài)和生理功能的調(diào)節(jié)上緊密相關(guān)。PS1除了影響線粒體的結(jié)構(gòu)和功能,也參與內(nèi)質(zhì)網(wǎng)的功能調(diào)節(jié)。我們前期也發(fā)現(xiàn)在PS1干擾的SH-SY5Y細胞模型上PS1缺失能夠引起內(nèi)質(zhì)網(wǎng)應(yīng)激,雖然應(yīng)激的強度較弱。然而,在PS1缺失情況下,我們對線粒體與內(nèi)質(zhì)網(wǎng)之間存在怎樣的作用關(guān)系,以及這種相互作用對細胞命運的影響并未了解很多。本研究發(fā)現(xiàn)PS1缺失引起的輕微內(nèi)質(zhì)網(wǎng)應(yīng)激對線粒體PARL-OPA1凋亡通路并不產(chǎn)生顯著影響,可當我們使用內(nèi)質(zhì)網(wǎng)應(yīng)激誘導(dǎo)劑在PS1干擾的SH-SY5Y細胞模型上誘導(dǎo)內(nèi)質(zhì)網(wǎng)輕度應(yīng)激時,線粒體PArL表達水平升高,線粒體內(nèi)細胞色素C (Cytochrome C)表達上升,但是在正常細胞上誘導(dǎo)輕度內(nèi)質(zhì)網(wǎng)應(yīng)激并沒有檢測出兩種蛋白的上調(diào),這提示我們,輕度內(nèi)質(zhì)網(wǎng)應(yīng)激可能激活了內(nèi)質(zhì)網(wǎng)調(diào)控細胞平衡的保護機制,使細胞更趨向于存活方向,由此抵消了PSl功能缺失所誘導(dǎo)的線粒體PARL-OPA1凋亡通路；同時,我們還發(fā)現(xiàn)這種保護作用不僅與內(nèi)質(zhì)網(wǎng)的應(yīng)激程度有關(guān),可能還具有PS1缺失的依賴性。因此,內(nèi)質(zhì)網(wǎng)應(yīng)激與線粒體之間相互作用的適度調(diào)節(jié)可能會成為抑制神經(jīng)細胞PS-PARL-OPA1凋亡通路激活的新途徑。在今后的研究中,不僅需要在PS 1/2 cDKO小、鼠上進一步驗證細胞模型上的結(jié)果,還需要深入探究“PS功能缺失”情況下線粒體與內(nèi)質(zhì)網(wǎng)的相互作用的確切分子調(diào)控機制。
[Abstract]:Alzheimer disease (ADD) is one of the most common neurodegenerative diseases.The neuronal fibrillary tangles caused by the accumulation of AP and the excessive phosphorylation of Tau protein were associated with the neuronal extracellular neuritis plaques in the brain. These pathological changes were considered to be the main reasons for the large number of apoptosis and cognitive impairment of neurocytes in the brain.Recent studies have shown that neuronal apoptosis is also associated with mitochondrial and endoplasmic reticulum dysfunction.We found functional and morphological abnormalities of neuronal mitochondria in the brain of PS1/PS2 conditioned double knockout PS1 / 2 conditional double knock out2 (PS1 / 2cDKO) mice, and in the SH-SY5Y cell model with PS1 interference (PS absence).We found that neuronal apoptosis induced by loss of PS1 function was regulated by mitochondrial PS1-PARL-OPA1 signaling pathway, that is, after over-expression of PARL in PS1 transient interference cell model, decreased OPA1 was up-regulated to normal level, and apoptosis was weakened.In this study, we further found that the expression of anti-apoptotic protein PARL and its regulated mitochondrial fusion protein OPA1 in PS1/2 cDKO mice were down-regulated at both RNA and protein levels, which was consistent with the results observed on SH-SY5Y cell model interfered with PS1.This study focused on the mitochondrial PS1-PARL-OPA1 apoptosis pathway. It was found that the overexpression of OPA1 did not change the low expression of PS1 and PARL after overexpression of OPA1 in the PS1 cell model, and the phenomenon of apoptosis did not change significantly, indicating that OPA1 was overexpressed but not shearing by PARL.Still unable to exercise its normal physiological function.As two independent organelles, mitochondria and endoplasmic reticulum Endoplasmic Reticulum.PS1 is closely related to the regulation of cellular homeostasis and physiological function. PS1 not only affects the structure and function of mitochondria, but also participates in the regulation of endoplasmic reticulum function.We also found that PS1 deletion induced endoplasmic reticulum stress in SH-SY5Y cell models with PS1 interference, although the stress intensity was weak.However, in the absence of PS1, we do not know much about the relationship between mitochondria and endoplasmic reticulum and the effect of this interaction on the fate of cells.In this study, we found that mild endoplasmic reticulum stress induced by PS1 deletion had no significant effect on mitochondrial PARL-OPA1 apoptosis pathway, but when we used endoplasmic reticulum stress inducer to induce mild endoplasmic reticulum stress on SH-SY5Y cell model with PS1 interference,Mitochondrial PArL expression increased, cytochrome C (cytochrome C) expression increased in mitochondria, but mild endoplasmic reticulum stress induced by mild endoplasmic reticulum stress in normal cells did not detect the up-regulation of two proteins.Mild endoplasmic reticulum stress may activate the protective mechanism of endoplasmic reticulum (ER) regulation of cell balance and make cells more likely to survive, thus counteracting the mitochondrial PARL-OPA1 apoptosis pathway induced by PSl dysfunction.We also found that this protective effect is not only related to the stress of endoplasmic reticulum, but also may be dependent on PS1 deletion.Therefore, the moderate regulation of the interaction between endoplasmic reticulum stress and mitochondria may be a new way to inhibit the activation of neuronal PS-PARL-OPA1 apoptosis pathway.In future studies, we need not only to further verify the results of the cell model in PS 1 / 2 cDKO mice, but also to explore the exact molecular regulation mechanism of the interaction between mitochondria and endoplasmic reticulum in the case of "PS functional deficiency".
【學位授予單位】：華東師范大學
【學位級別】：碩士
【學位授予年份】：2016
【分類號】：R749.16

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