P301L-Tau模型小鼠内嗅皮层-海马神经纤维变化与记忆功能障碍的相关性及其分子机制研究  

Correlation Analysis of the Entorhinal Cortex-Hippocampus Nerve Fiber Changes and Memory Dysfunction in P301L-Tau Mouse Model and Molecular Mechanism

作　　者：陈晓程 梁胜祥 林冰冰[1] 柳维林[2] 黄佳[1] CHEN Xiaocheng;LIANG Shengxiang;LIN Bingbing;LIU Weilin;HUANG Jia(College of Rehabilitation Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China;Rehabilitation Industry Institute,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学康复医学院,福建福州350122 [2]福建中医药大学康复产业研究院,福建福州350122

出　　处：《康复学报》2023年第2期136-141,共6页Rehabilitation Medicine

基　　金：国家自然科学基金项目(81774424);福建省教育厅高校“新世纪优秀人才支持计划”(闽教科〔2018〕47号)。

摘　　要：目的:探讨阿尔茨海默病(AD)P301L-Tau模型小鼠内嗅皮层-海马神经纤维改变与学习记忆功能的相关性及相关分子机制,为AD患者Tau蛋白靶向治疗提供依据。方法:选择16只3月龄雄性C57BL/6小鼠,按随机数字表法分为空载病毒对照组(对照组)和P301L-Tau模型组(模型组),每组8只。麻醉后,对照组小鼠双侧内嗅皮层注射空载病毒;模型组小鼠双侧内嗅皮层注射AAV-P301L-Tau病毒。注射完成后常规饲养至12月龄。采用Morris水迷宫测试评估小鼠学习记忆功能;采用9.4 T磁共振行弥散张量成像(DTI)扫描检测小鼠双侧内嗅皮层和海马的神经纤维变化情况;采用Western blot法检测小鼠内嗅皮层、海马t-Tau、p-Tau(Ser262)和糖元合成激酶-3β(GSK3β)蛋白的表达水平;采用Pearson相关性分析法分析小鼠内嗅皮层、海马各向异性分数(FA)与学习记忆功能的相关性。结果:①学习记忆功能测试:与对照组比较,模型组小鼠学习记忆功能明显下降,逃避潜伏期明显增加,穿越平台次数和目标象限时间占比明显降低(P<0.05)。②小鼠内嗅皮层和海马脑区中神经纤维结构:与对照组比较,模型组小鼠内嗅皮层和海马FA值均明显下降,内嗅皮层平均弥散率(MD)明显增加(P<0.05),而海马MD值差异无统计学意义(P>0.05)。③小鼠内嗅皮层、海马脑区FA值与学习记忆功能的相关性分析:2组小鼠内嗅皮层、海马FA值与穿越平台次数均呈正相关关系(r=0.76,P<0.05;r=0.62,P<0.05)。④小鼠内嗅皮层、海马脑区t-Tau、Ser262和GSK3β蛋白表达:与对照组比较,模型组内嗅皮层和海马t-Tau、p-Tau(Ser262)和GSK3β蛋白含量均明显增加(P<0.05)。结论:P301L-Tau模型小鼠内嗅皮层、海马脑区Tau蛋白过度磷酸化,导致小鼠内嗅皮层、海马脑区神经纤维结构遭受破坏和学习记忆功能障碍,其学习记忆功能障碍程度与内嗅皮层、海马神经纤维受损程度呈正相关。Objective:To observe the correlation between the changes of entorhinal cortex-hippocampus nerve fibers and learning and memory function in the P301L-Tau mouse model of Alzheimer's disease(AD)and related molecular mechanism,so as to provide the evidence for Tau-targeted treatment in AD patients.Methods:A total of 163-month-old male C57BL/6 mice were randomly divided into non-vector virus control group(control group)and P301L-Tau model group(model group),with eight mice in each group.After anesthesia,non-vector virus was injected into the bilateral entorhinal cortex of mice in the control group,and AAVP301L-Tau virus was injected into the bilateral entorhinal cortex of mice in the model group.After the injection,they were routinely fed to 12 months of age.Morris water maze test was used to detect the learning and memory function.Diffusion tensor imaging(DTI)scanning was used to detect the changes of nerve fibers in the bilateral entorhinal cortex and hippocampus.Western blot was used to detect the expression levels of t-Tau,p-Tau(Ser262),and glycogen synthase kinase-3β(GSK3β).Pearson correlation analysis was used to analyze the correlation between fractional anisotropy(FA)values in different brain regions of the entorhinal cortexhippocampus and their corresponding learning and memory functions.Results:(1)Learning and memory function test:compared with the control group,the learning and memory function of the model group significantly decreased,the escape latency significantly increased,the number of platform crossings and time ratio of target quadrant significantly decreased(P<0.05).(2)Structure of nerve fibers in the entorhinal cortex and hippocampus:compared with the control group,FA values of the entorhinal cortex and hippocampus in the model group decreased more significantly,MD values of the entorhinal cortex in the model group increased more significantly,while there was no significant difference in the MD values of the hippocampus in the model group(P>0.05).(3)Correlation analysis between FA values in the

关 键 词：阿尔茨海默病 磷酸化TAU蛋白 弥散张量成像 内嗅皮层-海马 学习记忆 

分 类 号：R749.16[医药卫生—神经病学与精神病学]