从小胶质细胞TLR4/NF-κB通路探讨针刺治疗缺血性中风的抗炎机制

项目名称： 从小胶质细胞TLR4/NF-κB通路探讨针刺治疗缺血性中风的抗炎机制

项目编号： No.81202765

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学八处

项目作者： 韩冰冰

作者单位： 山东中医药大学

项目金额： 23万元

中文摘要： 脑缺血免疫炎症反应促进了继发性脑损害，而中枢神经系统最主要的免疫效应细胞- - 小胶质细胞的活化则是脑缺血免疫炎症反应的关键始动因素。课题组以往的研究提示针刺可以减少脑缺血后小胶质细胞的活化，为深入探讨其机制，本项目拟建立MCAO大鼠模型，观察针刺对MCAO大鼠及TLR4阻断后MCAO大鼠的影响，采用神经行为学评分检测大鼠脑功能，HE及TUNEL染色检测神经元的坏死及凋亡，TTC染色检测梗塞灶面积，ELISA检测脑组织炎症因子含量，免疫荧光标记、激光共聚焦显微镜及原位RT-PCR检测小胶质细胞的活化、TLR4/NF-κB信号通路关键分子在小胶质细胞的表达。分析上述各因素之间的关系，论证"针刺可能通过影响小胶质细胞TLR4/NF-κB信号通路而调控小胶质细胞活化，抑制脑缺血免疫炎症损伤"的假说，本项目既是对以往研究的深入，也为阐明针刺治疗缺血性中风的脑保护机制提供新的研究思路和手段。

中文关键词： 针刺；免疫炎症；小胶质细胞；TLR4；

英文摘要： Immunoinflammatory response of cerebral ischemia can promote secondary cerebral injury. While the activation of microglia cells which are the main immune cells of central nervous system is the key initiating factor of immunoinflammatory response of cerebral ischemia. Our previous study indicated acupuncture could reduce the activation of microglia cells after cerebral ischemia. To investigate the mechanism, we will construct MCAO model rats and observe the effect of acupuncture on MCAO rats and the effect of acupuncture on MCAO rats after TLR4 is blocked. The brain function of MCAO rats will be detected by neurobehavioral score. Necrosis and apoptosis of nerve cells will be detected by HE and TUNEL staining. The area of infarct focus will be detected by TTC staining. The content of inflammatory factor in brain tissue will be detected by ELISA. The activation of microglia cells, the expressing of key molecular of TLR4/NF-κB signaling pathway on microglia cells will be detected by immune fluorescence labeling、laser scanning confocal microscope and RT-PCR. We will analyze the relation of the above factors, then discuss the presumption that "acupuncture can regulate the activation of microglia cells by influencing TLR4/NF-κB signaling path, finally restrain the immunoinflammatory damage of cerebral ischemia". This p

英文关键词： acupuncture；immune inflammation；microglia；TLR4；