基于平衡突触再生与突触删除研究针刺神庭、百会改善MCAO大鼠学习记忆的机制

Clinical acupuncture shenting and baihui to cure the stroke patients ,which could best improve the ability of learning and memory after ischemic stroke, but the mechanism is unknown.And our previous study had found that the ultrastructure of hippocampus neurons and synaptic plasticity after acupuncture were improved, but failed to fully describe the mechanism of the electronacupuncture to improve learning and memory. Member of translated 《eLife》 published in 2016 about the synaptic delete parts of the mechanism. Normal synaptic delete is very important for the formation of the neural circuits and maintain, mainly in experience dependent synaptic plasticity processes such as learning and memory, etc. Based on the study we speculated that acupuncture may balance on promoting synaptic regeneration and synaptic elimination to improve learning and memory. This study will be carried out as follows: using MCAO model rats,and acupuncture shenting and baihui. The effect of acupuncture on improving the volume of the infarct、glutamic metabolites and the functional connection of the rats by using the small animal MRI.The glucose metabolism of the brain regions surrounding of the MCAO rats after acupunctured Shenting and Baihui are detected by small animal PET,which is labeled 18F-FDG.Using the water maze experiment to test the ability of learning and memory of the MCAO rats,the results of which will show the correlation between the change of energy metabolism of the area around the ischemia.And then the synaptic ultrastructure will be observed by transmission electron microscope The expression of GABA、PSD95、SynapsinⅠ、CamkⅡ and other related genes and protein of the MCAO rats will be tested by RT-PCR Western- blot. Results would strong support that the mechanism of electroacupuncture improves cognitive impairment after stroke.And also the results would application and effect evaluation to provide for clinical,and would provide basis experimental for clinical application and experimental foundation.

课题组临床针刺神庭、百会改善缺血性脑卒中后学习记忆疗效显著；前期基础研究发现针刺后海马区神经元超微结构及突触再生情况都有所改善，但具体作用机制不明。段树民院士及其团队于2016年在《eLife》发表了关于突触删除影响学习记忆的部分机制，正常的突触删除对神经环路的形成和维持非常重要，主要发生在经验依赖的突触可塑性过程如学习和记忆等，由此判断出突触再生与突触删除关系密切。基于此设立针刺可能通过促进平衡突触再生和突触删除以改善学习记忆的科学假说。本课题拟以大鼠学习记忆能力、脑梗死面积、海马区功能连接、磁共振波谱、主要脑区葡萄糖代谢、海马区神经元突触超微结构、海马区神经元长时程增强/长时程抑制、GABA、PSD95、SynapsinⅠ、CamkⅡ等相关基因及蛋白表达情况为指标，从活体小动物影像、电生理及分子水平共同阐明针刺改善脑卒中后学习记忆障碍的作用机制，为临床推广针刺治疗脑卒中提供实验支持。

课题组围绕针刺可能通过促进平衡突触再生和突触删除以改善学习记忆的科学假说，开展一系列研究。建立缺血再灌注脑卒中大鼠模型，以针刺神庭、百会两个穴位，明确了针刺督脉上神庭、百会穴位能改善MCAO大鼠的学习记忆能力；能减轻脑梗死面积；海马区功能连接增强，海马区神经元突触超微结构显示神经元形态改善，突触在针刺后数量增加，突触间隙明显缩小。明确了海马区神经元长时程增强/长时程抑制的动态变化，显示电针的正反馈作用。进一步明确了电针在蛋白水平的调节作用，GABA、PSD95、SynapsinⅠ、CamkⅡ等相关基因及蛋白表达在电针后均有不同程度的上调，趋势一致，进一步明确了电针的作用。从活体小动物影像、电生理及分子水平共同阐明针刺改善脑卒中后学习记忆障碍的作用机制，为临床推广针刺治疗脑卒中提供实验支持。