督脉电针治疗AD双转基因小鼠不同脑区神经元凋亡作用机制的研究

As an important part of traditional Chinese medicine, acupuncture has attracted extensive attention of the medical profession in the world. Many clinical and experimental researches show that DU meridian electric acupuncture treatment has curative effect on Alzheimer's disease (AD). From earlier research we found that the du meridian electric acupuncture can reduce abnormal expression of Aβ and p-tau, and thus it can improve the AD mice’s cognitive ability and the loss of hippocampal neurons. However, the internal mechanism how the treatment works in inhibiting cell apoptosis and neuron loss is still not completely clear. New research shows that neurotoxicity factor produced by Aβ1-42 and p-tau can induce neuronal apoptosis through activating JNK signaling pathway. Therefore, by observing brain metabolism from the overall level using advanced molecular imaging micro-PET, observing neurons apoptosis using TUNEL, and by analyzing the relationship of the key signal transduction molecules of JNK signaling pathway upstream and downstream with modern molecular biology techniques such as western bolt and real-time PCR, this study propose to reveal the regulation of neuronal damage of DU meridian electric acupuncture treatment and its mechanism, and realize the quantization and visualization of its effect on resistance to neurons apoptosis and neurons repair, and provide scientific basis for clinical treatment of AD by acupuncture and moxibustion.

针灸疗法是传统中医学的重要组成部分，越来越受到国内外医学界的广泛关注。许多临床和实验研究表明，督脉电针治疗阿尔茨海默病（AD）疗效肯定。前期研究我们发现，督脉电针可以减少Aβ和p-tau的异常表达，从而改善AD小鼠认知能力和海马神经元的丢失。然而，督脉电针抑制细胞凋亡及神经元丢失的内在机制尚未完全清晰。最新研究显示，Aβ1-42和P-tau所产生的神经毒性因子，可通过激活JNK信号通路诱导神经凋亡。因此，本课题拟应用先进的分子影像学micro-PET技术从整体水平观察大脑物质代谢情况，结合TUNEL观测神经元凋亡情况，运用western bolt和real-time PCR等现代分子生物学技术分析JNK信号通路上下游关键信号转导分子的相互关系，旨在揭示督脉电针对AD神经元损伤的调节和它的作用机制，并实现其抗神经元凋亡，神细胞修复的定量化和可视化，为针灸临床治疗AD提供科学依据。

针灸疗法是我国传统中医药的重要组成部分，其疗效显著且有独特的作用优势，具有多靶点、多水平、多途径的整体调节能力。督脉贯脊入络脑，通督电针是以“督脉--脑”系统为物质基础，“通督”为手段，“启神”为目的，通过疏通督脉发挥醒脑启神、益智开窍的效用。本课题旨在探讨通督电针抗老年性痴呆神经凋亡和改善记忆力的作用机制。本研究以通督电针为手段，从与老年性痴呆发病机制密切相关的JNK 神经元凋亡通路和突触可塑性为切入点，运用以mico-PET为检测手段、免疫组化、免疫荧光染色、Western blot等多种现代方法，从定位、定性、定量等不同水平观察通督电针治疗后不同脑区神经元变化的情况，同时结合发挥关键作用的信号通路相关蛋白的变化，探讨通督电针治疗老年性痴呆的作用机制。我们研究发现，督脉电针可有效改善阿尔茨海默病的学习记忆障碍，调节突触可塑性，提高痴呆小鼠的空间记忆能力，从而减少痴呆小鼠大脑神经元的损伤。督脉电针联合药物阻断剂（SP600125）可明显抑制JNK通路上下游的关键蛋白表达，减少上游激酶MKK7的表达，抑制下游关键蛋白c-Jun和p-JNK的过表达来减少Aβ蛋白沉淀，减少caspase-3的表达，从而达到抗凋亡的作用；督脉电针也可通过提高蛋白激酶A（PKA)和突触后致密物-95（PSD-95）的表达，达到改善记忆力的作用。由此可以得出，督脉电针可以通过调节突触可塑性、JNK信号途径多靶点抑制Aβ蛋白沉淀，改善学习记忆能力，减少神经凋亡的作用。