老年性痴呆同病异证的实验研究

发布时间：2018-04-04 14:00

本文选题：老年性痴呆　切入点：同病异证　出处：《山西中医学院》2016年硕士论文

【摘要】：目的:本课题以AD的两个病证结合模型:病(老年性痴呆,简称AD)加证(肾阳虚、痰浊阻窍)为研究对象,以该病的六个主要发病机制,即Tau蛋白异常、淀粉样前体蛋白代谢失调、胆碱能系统紊乱、神经细胞凋亡、泛素-蛋白酶体通路(UPP)异常、免疫炎症反应等的相关系列指标:Tau蛋白、Aβ、Ach和Ach E、Bax和Bcl-2、Ub和E1、P65为研究内容,利用分子生物学技术手段,探索AD两个典型证候的生物学内涵,寻找AD“同病异证”在生物学机制方面的共性及特异性。方法:将13只6月龄SAMR1小鼠分为正常组,39只6月龄SAMP8小鼠随机平均分为3组,每组13只,分别为AD模型组(简称模型组)、肾阳虚证AD模型组(简称肾虚组)、痰浊阻窍证AD模型组(简称痰浊组)。肾虚组采用腹腔内注射氢化可的松的方法复制肾阳虚证AD模型;痰浊组采用饲喂6W高脂饲料的方法复制痰浊阻窍证AD模型。模型复制成功后,采集所需标本,分别从光镜(肝脏和肾脏组织)、电镜(海马)、生化指标(血液、海马和大脑皮层)等不同层面,寻找AD肾阳虚证与痰浊阻窍证在生物学机制方面的共性与特异性。结果:1.形态学结果小鼠肝脏、肾脏光镜及海马电镜结果显示,正常组小鼠的肝脏、肾脏及海马组织超微结构未见明显异常;模型组小鼠的肝脏、肾脏及海马组织超微结构有轻度损害;痰浊组小鼠各组织的损害较正常组略重,肾虚组小鼠各组织的损害最重。2.海马、大脑皮层和血液生化指标检测结果2.1模型组与正常组比较:两组小鼠大脑皮层中Tau蛋白、Aβ和海马中Bax含量有显著性差异(P0.05),模型组Tau蛋白、Aβ、Bax含量较正常组高。两组小鼠在大脑皮层中Ach和AchE以及海马中Bcl-2、Ub和E1、P65指标方面均无显著性差异。2.2肾虚组与正常组比较:两组小鼠血液中ACTH和CORT含量均有显著性差异(P0.05),肾虚组小鼠ACTH和CORT含量较正常组低。2.3痰浊组与正常组比较:两组小鼠血液中TG含量有显著性差异(P0.05),痰浊组TG含量较正常组高,且为正常组TG含量的1.56倍。2.4肾虚组与痰浊组比较:肾虚组小鼠大脑皮层中Tau蛋白、Aβ和海马中Bax含量较痰浊组高,海马中Bcl-2含量较痰浊组低,差异有统计学意义(P0.05)。两组小鼠大脑皮层中Ach和AchE及海马中Ub和E1、P65含量无统计学差异。结论:肾阳虚证与痰浊阻窍证时AD的以下三个机制,即Tau蛋白异常、淀粉样前体蛋白代谢失调和神经细胞凋亡会有明显的异常,肾阳虚证时此三种机制的异常较痰浊阻窍证时更明显,肾阳虚证时AD小鼠大脑的损害较痰浊阻窍证时的损害表现更突出。
[Abstract]:Objective: in this study, two models of AD combined with syndromes: disease (AD) plus syndrome (kidney-yang deficiency, phlegm-turbid obstruction of orifices) were used to study the six main pathogenesis of AD, that is, abnormal Tau protein.Amyloid precursor protein metabolism disorder, cholinergic system disorder, neuronal apoptosis, UPP abnormality of ubiquitin proteasome pathway, immune inflammatory response, and other related indexes, such as A 尾 -Ach, Ach eau Bax, Bcl-2Ub and E1P65, were studied.By means of molecular biology technique, the biological connotation of two typical symptoms of AD was explored, and the commonness and specificity of the biological mechanism of AD "same disease and different syndrome" were found.Methods: thirteen 6-month-old SAMR1 mice were divided into normal group (n = 39) and 6-month-old SAMP8 mice (n = 39).AD model group (abbreviated as model group), kidney yang deficiency syndrome AD model group (abbreviated as kidney deficiency group), phlegm turbid obstruction syndrome AD model group (abbreviated phlegm turbid group).The AD model of kidney-yang deficiency syndrome was induced by intraperitoneal injection of hydrocortisone in kidney-deficiency group, and the AD model of phlegm-turbid obstruction syndrome was induced by feeding 6W high-fat diet in phlegm-turbid group.After the model was successfully duplicated, samples were collected from different levels, such as light microscope (liver and kidney tissue, electron microscope (hippocampus), biochemical indexes (blood, hippocampus and cerebral cortex), etc.To find the commonness and specificity of AD kidney yang deficiency syndrome and phlegm turbid obstruction syndrome in biological mechanism.The result is 1: 1.The ultrastructure of liver, kidney and hippocampal tissue in normal group was not abnormal, while in model group there was slight damage in liver, kidney and hippocampal tissue.The damage of each tissue in phlegm turbid group was slightly more serious than that in normal group, and that in kidney deficiency group was the most serious. 2.Results the biochemical indexes of hippocampus, cerebral cortex and blood in the model group were compared with those in the normal group: there was a significant difference in the contents of Tau protein A 尾 and Bax in hippocampus between the two groups. The content of Tau protein A 尾 -Bax in the model group was higher than that in the normal group.Compared with the normal group, the content of CORT in the blood of the group with phlegm turbidity was significantly higher than that in the group of phlegm turbidity (P 0.05), and the content of TG in the group of phlegm turbidity was higher than that in the group of normal phlegm.Compared with phlegm turbid group, the content of Tau protein A 尾 in cerebral cortex and Bax in hippocampus in kidney deficiency group were higher than those in phlegm turbid group, and Bcl-2 content in hippocampus was lower than that in phlegm turbid group (P 0.05).There was no significant difference in the contents of Ach and AchE in cerebral cortex and Ub and E1p65 in hippocampus between the two groups.Conclusion: there are three mechanisms of AD in the syndrome of kidney-yang deficiency and phlegm-turbid obstruction of orifices, that is, abnormal Tau protein, abnormal metabolism of amyloid precursor protein and apoptosis of nerve cells.The abnormality of these three mechanisms in kidney yang deficiency syndrome was more obvious than that in phlegm turbid obstruction of orifices syndrome. The brain damage of AD mice with kidney yang deficiency syndrome was more prominent than that of phlegm turbid orifices syndrome.
【学位授予单位】：山西中医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R277.7

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