本文選題：Alzheimer病　切入點(diǎn)：STZ側(cè)腦室注射　出處：《山西醫(yī)科大學(xué)》2013年碩士論文

【摘要】：雖然已經(jīng)有五種FDA認(rèn)證的藥物（tacrine, rivastigmine,galantamine, donepezil, and memantine）可以有效的延緩Alzheimer�。ˋD）的發(fā)病進(jìn)程，但是至今依然缺乏對AD這種中樞神經(jīng)退行性疾病有效的治療策略。目前認(rèn)為，有效阻止AD的方法是依賴于早期的診斷和預(yù)防。對于中藥治療AD的研究不失為一種AD的早期預(yù)防與治療有效策略。 AD與2型糖尿病（T2DM）之間的密切關(guān)系目前已經(jīng)得到了廣泛的證實(shí)�；趦烧叩墓餐l(fā)病機(jī)制，糖尿病治療藥物或許可以發(fā)揮神經(jīng)保護(hù)作用。通過激活GLP-1受體，GLP-1類似物已經(jīng)成為了一種臨床上成熟的糖尿病治療新藥。同時，在動物實(shí)驗(yàn)中，GLP-1類似物則被證實(shí)具有對AD動物模型的神經(jīng)保護(hù)效應(yīng)。 本實(shí)驗(yàn)所采用的中藥單體京尼平苷（Geniposide）已被證明是一種GLP-1受體激動劑，同時在培養(yǎng)細(xì)胞水平已經(jīng)被證實(shí)具有神經(jīng)保護(hù)作用。本實(shí)驗(yàn)旨在從AD動物模型的水平證實(shí)京尼平苷的神經(jīng)保護(hù)效應(yīng)。采用鏈脲佐菌素（STZ）側(cè)腦室注射形成AD大鼠模型。通過Morris水迷宮實(shí)驗(yàn)和免疫組織化學(xué)實(shí)驗(yàn)我們證實(shí)了單次側(cè)腦室注射京尼平苷（50μM,10μl）有效的阻止了約40%的STZ誘導(dǎo)的空間認(rèn)知損傷和約30%的STZ誘導(dǎo)的tau蛋白的過度磷酸化。與我們的預(yù)期相反，在免疫組化試驗(yàn)中，我們并沒有在STZ誘導(dǎo)的大鼠腦內(nèi)觀察到AD的另一重要病理變化β-淀粉樣蛋白（Aβ）的聚集，同時westernblot測定也顯示Aβ的前體蛋白——淀粉樣前體蛋白（APP）在STZ大鼠腦內(nèi)的表達(dá)也沒有明顯升高。在治療組的基礎(chǔ)上，我們利用PI3K的特異性抑制劑wortmannin（WT）阻止PI3K活性從而引起其下游蛋白GSK3的去抑制，以此干預(yù)治療組從而探究PI3K/GSK3信號通路在京尼平苷神經(jīng)保護(hù)作用中扮演的角色。結(jié)果顯示，，WT一定程度上阻止了京尼平苷對STZ誘導(dǎo)的大鼠行為學(xué)保護(hù)作用以及tau蛋白過度磷酸化的緩解作用，這一結(jié)果證實(shí)了PI3K/GSK3在京尼平苷的神經(jīng)保護(hù)作用中扮演著關(guān)鍵角色。 GSK3是一種重要的參與tau蛋白磷酸化調(diào)節(jié)的激酶。GSK3的過度活化在AD的多種病理變化有著廣泛的相關(guān)性。通過westernblot實(shí)驗(yàn)，我們發(fā)現(xiàn)STZ側(cè)腦室注射引發(fā)了腦內(nèi)顳葉皮層GSK3β的過度活化。同時，京尼平苷通過對GSK3β兩個調(diào)控位點(diǎn)的調(diào)節(jié)——上調(diào)抑制位點(diǎn)Ser9的磷酸化，下調(diào)激活位點(diǎn)Tyr216的磷酸化——抑制了STZ誘導(dǎo)的GSK3β的過度活化。 最后，我們通過透射電鏡進(jìn)行超微結(jié)構(gòu)觀察并發(fā)現(xiàn)京尼平苷阻止了STZ誘導(dǎo)的超微結(jié)構(gòu)變化，包括雙螺旋細(xì)絲（PHFs）樣結(jié)構(gòu)，突觸前膜囊泡的過度聚集，內(nèi)質(zhì)網(wǎng)異常，以及以暗細(xì)胞為特征的早期凋亡�？傊�，我們的研究證明了京尼平苷對于STZ急性側(cè)腦室注射誘導(dǎo)的AD大鼠模型的神經(jīng)保護(hù)作用，這也提示京尼平苷可以作為一種AD預(yù)防或治療的新藥進(jìn)行進(jìn)一步的研究。
[Abstract]:Although there are already five FDA certified drugs, such as tacrine, rivastigmine galantamine, donepezil, and memantine, which can effectively delay the onset of Alzheimer, there is still a lack of effective treatment strategy for AD, a central nervous degenerative disease. The effective method of preventing AD depends on the early diagnosis and prevention. The research on the treatment of AD with traditional Chinese medicine is an effective strategy for early prevention and treatment of AD. The close relationship between AD and type 2 diabetes mellitus (T2DM) has been widely confirmed. Diabetes drugs may play a neuroprotective role. By activating the GLP-1 receptor GLP-1 analogue, it has become a clinically mature new drug for the treatment of diabetes. In animal experiments, GLP-1 analogues have been shown to have neuroprotective effects on AD animal models. Geniposide, a traditional Chinese medicine monomer, has been proved to be a GLP-1 receptor agonist. At the same time, neuroprotective effects of geniposide have been proved at the level of cultured cells. The purpose of this experiment was to confirm the neuroprotective effect of geniposide from the level of AD animal model. The rat model of AD was formed by injecting streptozotocin (STZ) into lateral ventricle. By Morris water maze test and immunohistochemistry, we have proved that a single intracerebroventricular injection of geniposide (50 渭 m ~ (10 渭 l)) can effectively prevent spatial cognitive impairment induced by about 40% STZ and tau protein hyperphosphorous induced by about 30% STZ. Acidification. Contrary to our expectations, In the immunohistochemical test, we did not observe another important pathological change of AD, 尾 -amyloid A 尾, in the brain of rats induced by STZ. Westernblot analysis also showed that the expression of amyloid precursor protein (app) of A 尾 in the brain of STZ rats was not significantly increased. We used the specific inhibitor of PI3K, wortmanningnan WTA, to block the activity of PI3K and induce the desuppression of its downstream protein GSK3. In this intervention group, the role of PI3K/GSK3 signaling pathway in the neuroprotection of genipin was explored. The results showed that WT prevented the behavioral protective effect of genipin on STZ induced rats and tau protein to some extent. The mitigation of hyperphosphorylation, The results confirm that PI3K/GSK3 plays a key role in the neuroprotective effect of genipin. The over-activation of GSK3, an important kinase involved in the phosphorylation of tau protein, has a broad correlation with various pathological changes in AD. We found that intracerebroventricular injection of STZ induced excessive activation of GSK3 尾 in the temporal cortex of the brain. At the same time, geniposide up-regulated the phosphorylation of Ser9, the inhibitory site, by regulating the two regulatory sites of GSK3 尾. Down-regulating the phosphorylation of Tyr216, the activation site, inhibited the overexpression of GSK3 尾 induced by STZ. Finally, we observed the ultrastructure of STZ by transmission electron microscopy and found that geniposide prevented the ultrastructural changes induced by STZ, including the hyperaggregation of presynaptic vesicles and abnormal endoplasmic reticulum (ER). In conclusion, our study demonstrated the neuroprotective effect of genipin on AD rat model induced by acute intracerebroventricular injection of STZ. This suggests that geniposide can be further studied as a new drug for AD prevention or treatment.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R749.1

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

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2 陳太忠;張艷;熊_";徐發(fā)良;龔建平;余正;;脂多糖預(yù)處理對GSK-3功能活性的影響[J];第三軍醫(yī)大學(xué)學(xué)報;2012年13期

3 范鳴s

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