本文選題：PPARγ + 羅格列酮�。� 參考：《寧波大學(xué)》2012年碩士論文

【摘要】：目的1.研究可溶性β淀粉樣蛋白寡聚體42（soluble amyloid-β protein42oligomers, SO Aβ42）處理對體外培養(yǎng)大鼠海馬神經(jīng)元樹突絲的密度、運動速度和長度以及樹突棘密度的潛在損傷效應(yīng)，以了解SO Aβ42對海馬神經(jīng)元突觸發(fā)育的影響。2.研究過氧化物酶體增殖物激活受體（peroxisome proliferator-activatedreceptor gamma, PPAR）激動劑羅格列酮預(yù)處理對SO Aβ介導(dǎo)的海馬神經(jīng)元突觸發(fā)育損傷的影響及其量效關(guān)系，并探討PPARγ在羅格列酮發(fā)揮功能中的作用。3.探尋羅格列酮在以上研究中發(fā)揮潛在作用的可能性機(jī)制。最終為進(jìn)一步了解阿爾茨海默氏�。ˋlzheimer’s disease, AD）的發(fā)病機(jī)制以及探尋AD的治療靶點和治療藥物提供實驗依據(jù)。 方法取新生第1天Wistar大鼠的海馬回做原代神經(jīng)元培養(yǎng)。在體外培養(yǎng)第5天（day5in vitro, DIV5），用定位在膜上的綠色熒光蛋白（F-GFP）和GFP標(biāo)記的肌動蛋白（GFP-actin）對神經(jīng)元進(jìn)行共轉(zhuǎn)染，以顯示神經(jīng)元樹突及其突起的詳細(xì)形態(tài)學(xué)細(xì)節(jié)；在DIV7和DIV15，實驗組給予神經(jīng)元500nM SO Aβ42孵育3h，干預(yù)組在給予SO Aβ42孵育前對神經(jīng)元分別給予0.1μM、0.5μM，5μM羅格列酮或5μM羅格列酮+5μM GW9662預(yù)孵育24h。通過活細(xì)胞成像方法觀察各試驗組DIV7神經(jīng)元樹突絲的密度、運動速度和長度以及DIV15神經(jīng)元樹突棘密度的情況。其次，在DIV5，用F-GFP、GFP-actin和靶向線粒體表達(dá)的紅色熒光蛋白（Mitochondrially targeted red fluorescent protein, MitoRed）對神經(jīng)元進(jìn)行共轉(zhuǎn)染，以顯示神經(jīng)元樹突和其突起以及神經(jīng)元線粒體的形態(tài)學(xué)細(xì)節(jié)；在DIV15，實驗組給予神經(jīng)元500nM SO Aβ42孵育3h，干預(yù)組在給予SO Aβ42孵育前對神經(jīng)元給予5μM羅格列酮預(yù)孵育24h，通過活細(xì)胞成像方法觀察相應(yīng)試驗組DIV15神經(jīng)元樹突中線粒體分布的情況。 結(jié)果1.單獨500nM SO Aβ42處理3h導(dǎo)致DIV7大鼠海馬神經(jīng)元樹突絲的密度明顯下降；羅格列酮預(yù)處理24h以劑量依賴的方式逆轉(zhuǎn)SO Aβ42介導(dǎo)的樹突絲密度減小。SO Aβ42和羅格列酮處理均不引起樹突絲的運動速度和長度發(fā)生改變。2.單獨500nM SO Aβ42處理3h引起DIV15大鼠海馬神經(jīng)元樹突棘密度顯著性減��；羅格列酮預(yù)處理24h以劑量依賴的方式逆轉(zhuǎn)SO Aβ42介導(dǎo)的樹突棘密度下降。3.5μM PPARγ特異性拮抗劑GW9662和5μM羅格列酮對神經(jīng)元共同預(yù)處理有效阻斷5μM羅格列酮逆轉(zhuǎn)SO Aβ42介導(dǎo)的神經(jīng)元樹突絲或樹突棘密度下降。4.單獨500nM SO Aβ42處理3h降低了DIV15大鼠海馬神經(jīng)元樹突中線粒體總長度與對應(yīng)樹突干總長度的比值、樹突中離散線粒體的密度以及含線粒體的樹突棘數(shù)目與總樹突棘數(shù)目的比例，SO Aβ42處理引起的該現(xiàn)象在給予神經(jīng)元5μM羅格列酮預(yù)處理24h后被有效逆轉(zhuǎn)。SO Aβ42和羅格列酮處理均不改變樹突中線粒體的長度。 結(jié)論SO Aβ42處理對體外培養(yǎng)大鼠海馬神經(jīng)元的突觸發(fā)育存在損傷作用，，導(dǎo)致神經(jīng)元的突觸數(shù)目下降；羅格列酮預(yù)處理可以濃度依賴地逆轉(zhuǎn)SO Aβ42介導(dǎo)的神經(jīng)元突觸發(fā)育損傷，該效應(yīng)通過羅格列酮激活PPARγ受體起作用。羅格列酮的保護(hù)效應(yīng)機(jī)制可能與其可以逆轉(zhuǎn)SO Aβ42介導(dǎo)的神經(jīng)元樹突中線粒體分布異常，包括樹突中線粒體數(shù)目減少以及含線粒體的樹突棘比例下降有關(guān)。
[Abstract]:Objective 1. to study the effects of soluble beta amyloid oligomer 42 (soluble amyloid- beta protein42oligomers, SO A beta 42) on the density, movement speed, length and dendrite density of dendritic spines in cultured rat hippocampal neurons in vitro, in order to understand the effect of SO A beta 42 on the synaptic development of hippocampal neurons by.2. peroxidation The effect of rosiglitazone preconditioning with peroxisome proliferator-activatedreceptor gamma (PPAR) agonist roglitazone on the synaptic development damage of hippocampal neurons mediated by SO A beta and its quantitative effect relationship, and to explore the potential of PPAR gamma in the function of rosiglitazone to explore the potential of rosiglitazone to play potential in the above study. The possible mechanism of the action can provide an experimental basis for further understanding the pathogenesis of Alzheimer 's disease (AD) and the target of the treatment of AD and the treatment of drugs.
Methods the primary cultured hippocampal neurons of Wistar rats were cultured for first days. The neurons were cultured in vitro for fifth days (day5in vitro, DIV5). The neurons were co transfected with the green fluorescent protein (F-GFP) and the GFP labeled actin (GFP-actin), in order to show the detailed morphological details of the neuron dendrites and their protrusions; in DIV7 and DIV15, the experimental group was incubated with 500nM SO A beta 42 to incubate 3h, and the intervention group was given 0.1 mu M, 0.5 micron, 5 u M rosiglitazone or 5 mu M rosiglitazone before incubating SO A beta. Second, in DIV5, the neurons were co transfected with F-GFP, GFP-actin and Mitochondrially targeted red fluorescent protein (MitoRed) expressed in the target mitochondria in order to show the morphological details of the neuron dendrites and its protuberances and the mitochondria of the divinity. In DIV15, the experimental group was given. The neurons were incubated with 500nM SO A beta 42 for 3h, and the intervention group was incubated with 5 u M rosiglitazone before SO A beta 42 incubated to incubate 24h, and the distribution of mitochondria in the dendrites of the experimental group was observed by living cell imaging.
Results 1. 500nM SO A beta 42 treated 3H resulted in a significant decrease in the density of the dendrites of the hippocampal neurons in DIV7 rats; rosiglitazone pretreated 24h in a dose-dependent manner to reverse the dendrite density mediated by SO A beta 42, which reduced the.SO A beta 42 and the rosiglitazone treatment without causing the change of the kinetic speed and length of the dendrites. The density of dendritic spines in hippocampal neurons of DIV15 rats was significantly reduced by 3H treatment with beta 42; rosiglitazone pretreated 24h in a dose dependent manner to reverse the density of dendritic spines mediated by SO A beta 42,.3.5 mu M PPAR gamma specific antagonist GW9662 and 5 micron M rosiglitazone, which effectively blocked 5 micron M rosiglitazone to reverse SO 42. The density of neuron dendrites or dendrites decreased.4. alone 500nM SO A beta 42 treatment 3H reduced the ratio of the total length of mitochondria to the total length of the dendrites of the hippocampal neurons of DIV15 rats, the density of the discrete mitochondria in the dendrites and the proportion of the number of dendritic spines with the mitochondrial dendrites and the number of the total dendritic spines, SO A beta 42 treatment caused the 3H This phenomenon was reversed effectively after giving neurons 5 micron M rosiglitazone pretreatment 24h..SO A A 42 and rosiglitazone did not change the length of mitochondria in dendrites.
Conclusion SO A beta 42 treatment can damage the synaptic development of hippocampal neurons in vitro, and lead to a decrease in the number of synapses. Rosiglitazone preconditioning can reverse the synaptic development damage induced by SO A beta 42. This effect is mediated by rosiglitazone activation of PPAR gamma receptor. The protection of rosiglitazone The effect mechanism may reverse the abnormal mitochondrial distribution in the neuron dendrites mediated by SO A beta 42, including the decrease in the number of mitochondria in the dendrites and the decrease in the proportion of the dendritic spines containing the mitochondria.
【學(xué)位授予單位】：寧波大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R749.16

【共引文獻(xiàn)】

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