金思维提取物对3×Tg-AD转基因小鼠脑内蛋白质质量控制网络的影响

Background: β-amyloid deposition and hyperphosphorylated tau are two core pathological changes in Alzheimer disease (AD), which have been found closely related with the dysregulation of protein quality control networks, thus the disruption of proteins' production, folding and degradation. Aim: GEPT, a Chinese herbal formula with activating blood and dissipating phlegm function, have been found effective in treating AD. Thus, present research aims at observing the influence of GEPT on behavioral, hippocampus and cortical pathological (ultra-structure of neurons and synapses, and β-amyloid and hyperphosphorylated tau depositions) changes in 3×Tg-AD transgenic mouse and then observing influence of GEPT on changes of chaperone, ubiquitin-proteasomal and autophagy- lysosomal systems in 3×Tg-AD mouse. Method: Diving platform experiment and Morris water maze experiment will be used to check memory and behavior of 3×Tg-AD transgenic mouse; electron microscope will be applied to observe ultra-structure of neurons; immunohistochemisty, Western blot and ELISA will be used to test expressions ofβ-amyloid tau and key proteins in chaperone system, ubiquitin proteasome system and autophagy-lysosome system. Application: The whole picture of protein quality control net in 3×Tg-AD transgenic mouse will be depicted and pharmaceutical targets in this networks of GEPT will be revealed.

背景：β-淀粉样蛋白(Aβ)和过度磷酸化Tau蛋白(P-Tau) 的堆积是阿尔茨海默病（AD）的主要病理改变，这与蛋白质质量控制网络异常及蛋白折叠异常和降解受阻有关。前期临床研究发现，金思维提取物补肾化痰能够显著改善AD患者的认知症状。目的：评价补肾化痰法中药对3×Tg-AD转基因小鼠脑内分子伴侣蛋白、泛素-蛋白酶体和自吞噬-溶酶体系统的影响。方法：应用跳台实验和Morris水迷宫检测小鼠空间记忆学习能力、电镜观察海马及皮层神经元、突触的超微结构，应用免疫组化、Western blot和ELISA法检测不同干预组Aβ、Tau、分子伴侣蛋白系统、泛素-蛋白酶体系统和自吞噬-溶酶体系统的关键蛋白的差异。意义：揭示补肾化痰法中药治疗AD的可能生物效应靶点。

β-淀粉样蛋白(Aβ)和过度磷酸化Tau 蛋白(P-Tau) 的堆积是阿尔茨海默病（AD）的主要病理改变，这与蛋白质质量控制网络异常及蛋白折叠异常和降解受阻有关。前期临床研究发现，金思维提取物（GEPT）补肾化痰能够显著改善AD 患者的认知症状，在此基础上，应用跳台实验和Morris 水迷宫检测小鼠空间记忆学习能力、电镜观察海马及皮层神经元、突触的超微结构，应用免疫组化检测不同干预组Aβ 、Tau、分子伴侣蛋白系统、泛素-蛋白酶体系统和自吞噬-溶酶体系统的关键蛋白的差异，发现GEPT能够降低3×Tg-AD转基因小鼠脑内β-淀粉样肽的表达，进而改善神经突触损伤与空间学习记忆能力；能够降低3×Tg-AD转基因小鼠脑内过度磷酸化的Tau的表达，防止神经纤维缠结形成带来的损害；可上调3×Tg-AD转基因小鼠脑内的分子伴侣蛋白系统的关键调节靶点HSP70；可上调3×Tg-AD转基因小鼠脑内的泛素-蛋白溶酶体系统关键调节靶点泛素蛋白，可调节3×Tg-AD转基因小鼠脑内的自吞噬-溶酶体系统关键调节靶点LC3B，提示GEPT对蛋白质质量控制网络存在干预作用，进而发挥神经保护作用，揭示了补肾化痰法中药治疗AD 的可能生物效应靶点。