家族型與散發(fā)型阿爾茨海默病動(dòng)物模型的對(duì)比分析
發(fā)布時(shí)間：2019-02-27 17:24
【摘要】：目的：隨著我國(guó)老齡人口數(shù)量的日益增多，阿爾茨海默病（Alzheimer’s disease，AD)這種主要在老年群體中出現(xiàn)的疾病,將會(huì)給社會(huì)和家庭帶來極大危害。AD分為散發(fā)型（SAD）和家族型（FAD）。大部分AD患者為散發(fā)型，病因可能來自多個(gè)因素，包括環(huán)境，基因和代謝等。家族型是由于早老素或是APP的突變引起。實(shí)驗(yàn)中最常應(yīng)用的AD鼠模型是三轉(zhuǎn)基因鼠模型（3×Tg-AD），它可以使大腦中出現(xiàn)突變的早老素1，APP和過表達(dá)的磷酸化tau蛋白，因此代表了家族型AD鼠模型。散發(fā)型AD鼠模型可以通過往小鼠腦室內(nèi)注射鏈霉素（STZ）實(shí)現(xiàn)。盡管這兩類模型已經(jīng)廣泛引用于科研中，但是兩者之間的差異并沒有被報(bào)道。我們實(shí)驗(yàn)的目的是從APP形成/tau蛋白，突觸功能，凋亡和自嗜，AD相關(guān)蛋白激酶，葡萄糖代謝，胰島素信號(hào)、mTOR通路以及炎癥等八個(gè)方面，篩選出不同類型的AD鼠模型參與其分子生物學(xué)機(jī)制的基因和蛋白表達(dá)的異同，對(duì)AD的研究和藥物的發(fā)展有十分重要的社會(huì)意義和不可估量的經(jīng)濟(jì)價(jià)值。 方法：我們應(yīng)用基因芯片技術(shù)，比較不同類型中小鼠的海馬和皮質(zhì)中84個(gè)AD相關(guān)基因的表達(dá)情況。這些基因涉及APP形成/tau蛋白，突觸功能，，凋亡和自嗜，AD相關(guān)蛋白激酶，葡萄糖代謝，胰島素信號(hào)以及mTOR通路等。本研究應(yīng)用PCR芯片數(shù)據(jù)分析系統(tǒng)處理研究不同類型AD鼠基因的表達(dá)差異，同時(shí)應(yīng)用蛋白印跡技術(shù)觀察小鼠腦組織炎癥改變情況，進(jìn)一步分析不同模型之間蛋白表達(dá)差異。我們還將冰凍的小鼠腦組織切片，進(jìn)行免疫組化染色，通過光鏡觀察病理情況，比較不同區(qū)域腦組織與蛋白表達(dá)的關(guān)系，對(duì)蛋白印跡的結(jié)果進(jìn)行驗(yàn)證。 結(jié)果：我們發(fā)現(xiàn)在兩種小鼠模型中，大約有20個(gè)來自上面分類中的基因有差異性表達(dá)。一些基因的改變和先前在AD大腦里面觀察結(jié)果是一致的。在兩種小鼠模型中，這些差異性基因的表達(dá)是減少或者是趨于下降的。兩種小鼠模型的大腦皮質(zhì)中基因表達(dá)的多樣性更加明顯。在三轉(zhuǎn)基因鼠中，更多的相關(guān)突觸功能的基因是下調(diào)的，反之，在STZ注射鼠中，更多的相關(guān)胰島素信號(hào)和葡萄糖代謝的基因是下調(diào)的。蛋白印跡技術(shù)可見兩組小鼠大腦中都有不同程度炎癥發(fā)生，存在蛋白差異表達(dá)。STZ注射鼠中炎癥發(fā)生的程度比三轉(zhuǎn)基因小鼠多。免疫組化技術(shù)進(jìn)行驗(yàn)證，發(fā)現(xiàn)蛋白表達(dá)情況和以上結(jié)果一致，并且發(fā)現(xiàn)兩組小鼠中海馬區(qū)域的炎癥反應(yīng)要高于其他區(qū)域。 結(jié)論：本課題闡述了STZ注射鼠和三轉(zhuǎn)基因鼠之間基因表達(dá)的相同和不同處，也為兩種模型中AD相關(guān)基因表達(dá)的改變提供了詳細(xì)的介紹。這種高通量檢測(cè)基因的實(shí)驗(yàn)?zāi)Ｊ�，為AD在基因水平的研究奠定了基礎(chǔ)。同時(shí)聯(lián)合蛋白印跡技術(shù)和免疫組化等多項(xiàng)實(shí)驗(yàn)技術(shù)，通過APP形成，tau蛋白/細(xì)胞骨架，突觸功能，凋亡和自嗜，AD相關(guān)蛋白激酶，葡萄糖代謝，胰島素信號(hào)、mTOR通路及炎癥等多方面的研究，篩選出參與不同類型AD鼠模型分子生物學(xué)機(jī)制的基因和蛋白的異同，為阿爾茨海默病的研究提供了新的思路，對(duì)AD的研究和藥物的發(fā)展有重大意義。
[Abstract]:Objective: With the increasing number of aging population in our country, the disease of Alzheimer's disease (AD), which is mainly in the elderly population, will bring great harm to the society and the family. AD is divided into an emission type (SAD) and a family type (FAD). Most of the AD patients are sporadic, and the cause may come from multiple factors, including the environment, gene and metabolism, and so on. The family type is caused by an early age or a mutation in the APP. The most commonly used model of AD mice in the experiment is a three-transgenic mouse model (3-GTg-AD), which can cause a mutation in the brain of 1, APP, and overexpressed phosphorylated tau proteins, thus representing a family-type AD mouse model. The sporadic AD mouse model can be achieved by injecting streptomycin (STZ) into the mouse brain. Although the two models have been widely cited in scientific research, the difference between the two models is not reported. The purpose of our experiment is to form the/ tau protein, the synaptic function, the apoptosis and the self-, AD-related protein kinase, the glucose metabolism, the insulin signal, the mTOR pathway, and the inflammation in eight aspects, The similarities and differences of gene and protein expression of different type of AD mouse model in molecular biological mechanism were selected, and the research of AD and the development of drug were of great social significance and inestimable economic value. Methods: We applied gene chip technique to compare the expression of 84 AD-related genes in the hippocampus and cortex of different types of mice. Conditions. These genes involved APP-forming/ tau protein, synaptic function, apoptosis and self-, AD-related protein kinase, glucose metabolism, insulin signal, and mTOR pathway In this study, the difference of expression of different types of AD mice was studied by PCR chip data analysis system, and the expression of protein in different models was further analyzed by using the Western blot technique. In addition, the frozen mouse brain tissue was cut, the immunohistochemical staining was carried out, the pathological condition was observed by light microscope, the relationship between the expression of brain and protein in different regions was compared, and the results of the Western blot were tested. The results: We found that in two mouse models, there were about 20 genes from the above classification. Sex expression. The changes in some genes and the previous observations in the AD brain are In two mouse models, the expression of these difference genes is reduced or tends to Down. The diversity of gene expression in the cerebral cortex of the two mouse models It is obvious that in the three transgenic mice, more of the genes associated with the synaptic function are down-regulated, whereas in the STZ mice, more of the genes associated with the insulin signal and the glucose metabolism are Down-regulation. Western blot showed that there were different levels of inflammation in the brain of both groups, and there was a protein difference. The degree of inflammation in the mice with STZ injection is higher than that of the three transgenes The expression of the protein was found to be consistent with the above results, and the inflammatory response of the hippocampus in the two groups was found to be higher than that of the two groups. Conclusion: This topic describes the same and different positions of the gene expression between the STZ injection and the three transgenic mice, and also provides for the change of the expression of the AD-related genes in the two models. The experimental mode of this high-throughput detection gene is the study of the gene level of AD. It has laid a foundation for many experiments, such as APP formation, tau protein/ cytoskeleton, synaptic function, apoptosis and self-, AD-related protein kinase, glucose metabolism, insulin signal, mTOR pathway and inflammation. In this study, the similarities and differences of the genes and proteins involved in the molecular biological mechanism of different types of AD mice were selected, and a new thought was provided for the study of Alzheimer's disease, and the research of AD and the development of drugs were provided.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R749.16;R-332
本文編號(hào)：2431462
[Abstract]:Objective: With the increasing number of aging population in our country, the disease of Alzheimer's disease (AD), which is mainly in the elderly population, will bring great harm to the society and the family. AD is divided into an emission type (SAD) and a family type (FAD). Most of the AD patients are sporadic, and the cause may come from multiple factors, including the environment, gene and metabolism, and so on. The family type is caused by an early age or a mutation in the APP. The most commonly used model of AD mice in the experiment is a three-transgenic mouse model (3-GTg-AD), which can cause a mutation in the brain of 1, APP, and overexpressed phosphorylated tau proteins, thus representing a family-type AD mouse model. The sporadic AD mouse model can be achieved by injecting streptomycin (STZ) into the mouse brain. Although the two models have been widely cited in scientific research, the difference between the two models is not reported. The purpose of our experiment is to form the/ tau protein, the synaptic function, the apoptosis and the self-, AD-related protein kinase, the glucose metabolism, the insulin signal, the mTOR pathway, and the inflammation in eight aspects, The similarities and differences of gene and protein expression of different type of AD mouse model in molecular biological mechanism were selected, and the research of AD and the development of drug were of great social significance and inestimable economic value. Methods: We applied gene chip technique to compare the expression of 84 AD-related genes in the hippocampus and cortex of different types of mice. Conditions. These genes involved APP-forming/ tau protein, synaptic function, apoptosis and self-, AD-related protein kinase, glucose metabolism, insulin signal, and mTOR pathway In this study, the difference of expression of different types of AD mice was studied by PCR chip data analysis system, and the expression of protein in different models was further analyzed by using the Western blot technique. In addition, the frozen mouse brain tissue was cut, the immunohistochemical staining was carried out, the pathological condition was observed by light microscope, the relationship between the expression of brain and protein in different regions was compared, and the results of the Western blot were tested. The results: We found that in two mouse models, there were about 20 genes from the above classification. Sex expression. The changes in some genes and the previous observations in the AD brain are In two mouse models, the expression of these difference genes is reduced or tends to Down. The diversity of gene expression in the cerebral cortex of the two mouse models It is obvious that in the three transgenic mice, more of the genes associated with the synaptic function are down-regulated, whereas in the STZ mice, more of the genes associated with the insulin signal and the glucose metabolism are Down-regulation. Western blot showed that there were different levels of inflammation in the brain of both groups, and there was a protein difference. The degree of inflammation in the mice with STZ injection is higher than that of the three transgenes The expression of the protein was found to be consistent with the above results, and the inflammatory response of the hippocampus in the two groups was found to be higher than that of the two groups. Conclusion: This topic describes the same and different positions of the gene expression between the STZ injection and the three transgenic mice, and also provides for the change of the expression of the AD-related genes in the two models. The experimental mode of this high-throughput detection gene is the study of the gene level of AD. It has laid a foundation for many experiments, such as APP formation, tau protein/ cytoskeleton, synaptic function, apoptosis and self-, AD-related protein kinase, glucose metabolism, insulin signal, mTOR pathway and inflammation. In this study, the similarities and differences of the genes and proteins involved in the molecular biological mechanism of different types of AD mice were selected, and a new thought was provided for the study of Alzheimer's disease, and the research of AD and the development of drugs were provided.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類號(hào)】：R749.16;R-332
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