炎性反应在阿尔茨海默病认知功能障碍发生与发展中的作用

发布时间：2019-06-02 00:02

【摘要】：研究背景和目的：现有的研究表明，，阿尔茨海默病（AD）患者脑内淀粉样蛋白Aβ）沉积增多，炎性反应显著升高，然而，迄今为止，对于不同病理形式的AβAD脑内炎性反应之间的关系以及AD脑内炎性反应与认知功能损害之间的关系缺乏深入研究。本项研究在前期试验的基础上，深入探讨不同病理形式的Aβ与D脑内炎性反应之间的关系，以及AD脑内炎性反应与认知功能损害之间的关系，AD的防治提供新的思路。研究方法：以3.5月龄、6月龄和12月龄APPswe/PS1dE9双转基因AD小鼠及年龄相匹配的野生型（WT）C57BL/6小鼠为研究对象，通过Morris水迷宫实验检各组实验动物获得性空间学习能力和空间记忆能力，通过免疫组织化学和荧光染方法检测各组实验动物脑组织中Aβ淀粉样斑块、活化星形胶质细胞和活化小胶质胞的数量变化情况，通过ELISA方法检测各组实验动物脑组织中可溶性Aβ、不可性Aβ及炎性反应介质IL-1β、IL-6、TNFα和PGE2的表达变化情况。进而使用相关性分析方法，探讨不同病理形式的Aβ（总的Aβ斑块、纤维性Aβ斑块、可溶性Aβ和不可溶性Aβ）与AD脑内炎性反应之间的关系，以及AD脑内炎性反应与认知功能损害之间的关系。研究结果：（1）Morris水迷宫实验表明，3.5月龄APPswe/PS1dE9小鼠与对照组WT小鼠相比，在获得性训练阶段潜伏逃逸时间没有显著差异，6月龄和12月龄APPswe/PS1dE9小鼠与相对照的WT小鼠相比，潜伏逃逸期均显著延长。（2）免疫组织化学和荧光染色表明，3.5月龄组APPswe/PS1dE9小鼠脑内没有Aβ淀粉样斑块沉积，12月龄APPswe/PS1dE9小鼠与6月龄相比，总的Aβ斑块和纤维性Aβ斑块沉积均显著增加。进一步的ELISA实验显示，不同年龄组APPswe/PS1dE9小鼠脑内可溶性Aβ含量具有显著的组间差异，3.5月龄APPswe/PS1dE9小鼠脑内没有检测到不可溶性Aβ，而12月龄APPswe/PS1dE9小鼠与6月龄相比，脑内不可溶性Aβ含量显著增加。（3）免疫组织化学染色表明，与对照组WT小鼠相比，3.5月龄组的APPswe/PS1dE9小鼠脑内活化星形胶质细胞数量没有显著差异，6月龄组和12月龄组的APPswe/PS1dE9小鼠脑内活化星形胶质细胞的数量显著增加；不同年龄组APPswe/PS1dE9小鼠脑内活化星形胶质细胞和活化小胶质细胞数量具有显著的组间差异。进一步的ELISA实验显示，与对照组WT小鼠相比，不同年龄组APPswe/PS1dE9小鼠脑内IL-1β、IL-6、TNFα和PGE2含量均显著增加；不同年龄组APPswe/PS1dE9小鼠脑内IL-1β、IL-6、TNFα和PGE2具有显著的组间差异。（4）不同病理形式的Aβ（总的Aβ斑块、纤维性Aβ斑块、可溶性Aβ和不可溶性Aβ）与AD脑内炎性反应的相关性分析显示，不同月龄APPswe/PS1dE9小鼠脑内可溶性Aβ水平与活化胶质细胞、IL-1β、IL-6、TNFα和PGE2含量均呈显著相关，而在早期和晚期斑块阶段，总的Aβ斑块、纤维性Aβ斑块和不可溶性Aβ含量与炎性反应指标均无显著相关性。（5）炎性反应与认知功能损害之间相关性分析显示，3.5月龄APPswe/PS1dE9小鼠空间记忆能力与活化的小胶质细胞、IL-1β、IL-6、TNFα和PGE2的水平呈显著负相关；6月龄和12月龄APPswe/PS1dE9小鼠空间学习和记忆能力与活化的星形胶质细胞、活化的小胶质细胞、IL-1β、IL-6、TNFα和PGE2的水平呈显著负相关。研究结论：（1）APPswe/PS1dE9小鼠脑组织中炎性反应增强可能早在前体斑块阶段已经发生，在早期和晚期斑块阶段显著增加。（2）APPswe/PS1dE9小鼠脑组织中多种炎性标志物与可溶性Aβ的水平显著相关，而与Aβ淀粉样斑块或不可溶性Aβ无关。（3）APPswe/PS1dE9小鼠脑内多种炎性标志物与其空间学习和记忆能力显著相关。上述研究结果表明，可溶性Aβ可能是导致AD脑内炎性反应的主要因素，而脑内多种炎性途径可能进一步促进AD认知功能损害的发生和发展，提示针对多种炎症反应途径的抗炎治疗可能为AD的防治提供新的思路。
[Abstract]:Background and purpose of the study: The existing studies have shown that the deposition of amyloid A-like protein in the brain of the patients with Alzheimer's disease (AD) has increased and the inflammatory response is significantly increased, however, to date, The relationship between the inflammatory response of A-AD in different pathological forms and the relationship between the inflammatory response of AD and the impairment of cognitive function are not in-depth study. The relationship between A-type and D-brain inflammatory reaction in different pathological forms, as well as the relationship between the response of AD and the impairment of cognitive function, and the prevention and treatment of AD were discussed in this study. Methods: The acquired space learning ability and spatial memory of the experimental animals were tested by Morris water maze test in the study of the three-transgenic AD mice and the age-matched wild-type (WT) C57BL/6 mice at the age of 3.5 months,6 months and 12 months of age. The changes of A-type starch-like plaque, activated astrocytes and activated microglia in the brain of each group were detected by immunohistochemistry and fluorescence staining. The soluble A in the brain of each group of experimental animals was detected by ELISA. Expression of IL-1, IL-6, TNF-and PGE _ 2 in A-and inflammatory-response media In this paper, the relationship between A-type (total A-plaque, fibrous A-plaque, soluble A-and non-soluble A-type) and AD-brain inflammatory reaction in different pathological forms, as well as the correlation between the inflammatory reaction of AD and the impairment of cognitive function were discussed using the correlation analysis method. The results of the study were as follows: (1) The Morris water maze test showed that the latent escape time of APPswe/ PS1dE9 mice at the age of 3.5 months was not significantly different from that of the control group WT mice, and the latent escape period in the 6-and 12-month-old APPswe/ PS1dE9 mice were significantly different from those of the corresponding WT mice. The results showed that APPswe/ PS1dE9 mice in the 3.5-month-old group did not have A-type amyloid plaque deposit in the brain of the 3.5-month-old group. There was no significant difference in the content of soluble A in the brain of APPswe/ PS1dE9 in the age group, and no soluble A content was detected in the brain of APPswe/ PS1dE9 in the 3-month-old age group, while the content of soluble A-A in the brain of APPswe/ PS1dE9 in the 12-month-old group was significantly higher than that of the 6-month-old. There was no significant difference in the number of activated astrocytes in the brain of APPswe/ PS1dE9 mice in the 3.5-month-old group compared with the control group WT mice, and the number of activated astrocytes in the brain of the APPswe/ PS1dE9 mice in the 6-month-old group and the 12-month-old group was significantly higher than that of the control group WT mice. Increased number of activated astrocytes and activated microglia in the brain of APPswe/ PS1dE9 mice in different age groups The results showed that the levels of IL-1, IL-6, TNF, and PGE2 in the brain of the mice were significantly higher than those of the control group, and the levels of IL-1, IL-6, TNF and PGE2 in the brains of the mice were significantly higher than those in the control group. The correlation between the levels of soluble A in the brain of APPswe/ PS1dE9 mice and the activation of glial cells, IL-1, I, and I in the brain of APPswe/ PS1dE9 in different months of age showed that the level of soluble A-antigen in the brain of APPswe/ PS1dE9 in different months was the same as that of the activated glial cells. The levels of L-6, TNF and PGE2 were significantly correlated, while in the early and late stage of the plaque, the total A-A plaque, the fibrous A-plaque and the non-soluble A-A content and the inflammatory response index were not significant. The correlation between (5) inflammatory response and cognitive impairment showed that the spatial memory capacity of the 3.5-month-old APPswe/ PS1dE9 mice was significantly higher than that of activated microglia, IL-1, IL-6, TNF-and PGE2. The spatial learning and memory capacity of APPswe/ PS1dE9 in 6-and 12-month-old mice were significantly correlated with the activated astrocytes, activated microglia, IL-1, IL-6, TNF-and PGE2 levels. Conclusion: (1) The enhancement of inflammatory response in the brain of APPswe/ PS1dE9 mice may have occurred earlier in the stage of the precursor plaque, in the early and late stage of plaque formation. (2) The levels of multiple inflammatory markers in the brain tissue of the APPswe/ PS1dE9 mice were significantly correlated with the level of soluble A-type, and were similar to that of the A-type amyloid plaques or insolubles. (3) Multiple inflammatory markers in the brain of APPswe/ PS1dE9 mice and their spatial learning and memory The results of the above-mentioned study show that the soluble A-factor may be a major factor in the response of AD to the internal inflammatory response, while a wide range of inflammatory pathways in the brain may further contribute to the impairment of the AD cognitive function. Occurrence and development, suggesting that anti-inflammatory therapy for multiple inflammatory response pathways may be the prevention and treatment of AD
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R749.16

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