癫痫清颗粒对阿尔茨海默病模型小鼠Tau蛋白磷酸化及Aβ产生的影响

发布时间：2018-04-11 12:01

本文选题：癫痫清颗粒 + 阿尔茨海默病　；参考：《辽宁中医药大学》2017年硕士论文

【摘要】：目的:研究癫痫清颗粒对侧脑室注射Aβ_(1-42)所致阿尔茨海默病(AD)模型小鼠tau蛋白异常磷酸化和Aβ产生的影响,并探讨其相关分子机制。材料与方法:选取8周龄的昆明种小鼠(18-22 g),按体重随机分为六组,分别是假手术组,模型组,盐酸多奈哌齐1.3 mg/kg对照组,癫痫清颗粒12.48 g/kg、6.24 g/kg、3.12 g/kg剂量组。利用脑立体定位仪,向各组小鼠侧脑室一次性注射Aβ_(1-42)(1 mg/ml)3 ul造模(假手术组侧脑室注射等体积生理盐水)。于造模后第二天按每公斤体重给药量20 ml的剂量灌胃给药。连续给药21 d,每日一次。第15 d进行Y迷宫行为学实验,第16 d至21 d进行水迷宫行为学实验。考察癫痫清颗粒对AD模型小鼠行为学的影响。实验结束后取材,分别采用HE染色技术观察癫痫清颗粒对AD模型小鼠脑组织神经元细胞的影响;Western blot方法检测tau-5、Ser404、Thr181、GSK-3β、phospho-GSK-3β、Akt、phospho-Akt、PI3K、phospho-PI3K的表达,考察癫痫清颗粒对AD模型小鼠tau蛋白磷酸化的影响;采用免疫组织化学染色法观察Aβ_(1-42)、Aβ1-40在AD模型小鼠脑组织中的表达,ELISA法测定Aβ_(1-42)、Aβ1-40在AD模型小鼠脑组织中的含量,Western blot方法检测BACE1、PS1的表达,考察癫痫清颗粒对AD模型小鼠Aβ产生的影响。结果:1.癫痫清颗粒可以保护AD模型小鼠脑组织神经元细胞,提高AD模型小鼠的学习记忆能力;2.癫痫清颗粒可以降低Ser404、Thr181的蛋白表达,抑制tau蛋白异常磷酸化;3.癫痫清颗粒可以增加phospho-GSK-3β的蛋白表达,抑制GSK-3β的活性;4.癫痫清颗粒可以增加phospho-Akt/AKT、phospho-PI3K/PI3K的蛋白表达,激活PI3K/AKT信号通路;5.癫痫清颗粒可以降低Aβ_(1-42)、Aβ1-40在AD模型小鼠脑组织中的沉积,降低Aβ_(1-42)、Aβ1-40在AD模型小鼠脑组织中的含量;6.癫痫清颗粒可以降低BACE1、PS1的活性,从而降低Aβ的生成。结论:1.癫痫清颗粒可减轻Aβ_(1-42)引起AD模型小鼠神经元损伤,改善学习记忆障碍;2.癫痫清颗粒可能通过PI3K/Akt/GSK-3β信号通路,抑制tau蛋白异常磷酸化,延缓AD病程;3.癫痫清颗粒能通过抑制BACE1及PS1的活性,减少AD模型小鼠脑内Aβ含量,降低其对脑组织的损伤。
[Abstract]:Aim: to study the effect of tau protein abnormal phosphorylation and A 尾 production in AD mice induced by contralateral intracerebroventricular injection of Jiangqing granule (A 尾 1-42), and to explore the molecular mechanism of the abnormal phosphorylation and A 尾 production in mice with Alzheimer's disease (AD).Using the brain stereotactic locator, the model was established by single injection of A 尾 1-42 mg/ml)3 ul into the lateral ventricle of each group (sham operation group was injected with normal saline in the lateral ventricle of the same volume).On the second day after modeling, 20 ml / kg body weight was administered intragastrically.The drug was given once a day for 21 days.The behavior of Y maze and water maze were tested on 15th day and 16th to 21st day respectively.To investigate the effect of Qianqing granule on the behavior of AD model mice.Immunohistochemical staining method was used to observe the expression of A 尾 -Stap-1-42A1-40 in the brain of AD model mice. The content of A 尾 1-40 in brain tissue of AD model mice was determined by Elisa. Western blot method was used to detect the expression of BACE1-PS1, and the effect of Xianqing granule on A 尾 production in AD model mice was investigated.The result is 1: 1.Xianqing granule can protect brain neuron cells of AD model mice and improve learning and memory ability of AD model mice.Xianqing granule can reduce the protein expression of Ser404 and Thr181, and inhibit abnormal phosphorylation of tau protein.Qianqing granule can increase the protein expression of phospho-GSK-3 尾 and inhibit the activity of GSK-3 尾.Qianqing granule can increase the protein expression of phospho-Akt / AKTirphospho-PI3K / PI3K and activate the PI3K/AKT signaling pathway.Xianqing granule can reduce the deposition of A 尾 1-40 in the brain tissue of AD mice and the content of A 尾 1-40 in the brain tissue of AD model mice.Qianqing granule can reduce the activity of BACE1 and reduce the formation of A 尾.Conclusion 1.Xianqing granule can reduce neuron damage and improve learning and memory impairment in AD mice induced by A 尾 -tir 1-42).Jiangqing granule may inhibit the abnormal phosphorylation of tau protein and delay the course of AD by PI3K/Akt/GSK-3 尾 signaling pathway.Xianqing granule can reduce the content of A 尾 in the brain of AD model mice by inhibiting the activity of BACE1 and PS1, and decrease the damage to brain tissue.
【学位授予单位】：辽宁中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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