ATP经P2X4受体介导在全脑缺血再灌注损伤中的作用及机制的研究

项目名称： ATP经P2X4受体介导在全脑缺血再灌注损伤中的作用及机制的研究

项目编号： No.81471260

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 向正华

作者单位： 中国人民解放军第二军医大学

项目金额： 70万元

中文摘要： 缺血性脑中风是目前严重威胁人类健康的常见疾病。治疗原则是尽快恢复缺血区血供。然而缺血后再恢复血供往往会造成神经元的进一步损伤即脑缺血再灌注损伤（CIRI)。CIRI形成机制极复杂，文献提示可能与细胞外毒性物质(兴奋性氨基酸、细胞外ATP)聚集、细胞内钙离子超载、自由基生成等因素有关。动物实验发现全脑缺血再灌注后会诱发海马、大脑皮质和纹状体等区域发生CIRI发生。我们发现缺血再灌注后48小时正在凋亡的海马CA1区神经元高表达P2X4受体；P2X4受体拮抗剂可部分抑制神经元凋亡并改善损伤后动物的学习记忆功能；病毒载体使海马CA1神经元过表达P2X4受体后神经元出现凋亡。上述预实验清楚地说明海马CA1的CIRI与P2X4受体上调相关。本实验将通过组织微透析、蛋白印迹、钙离子测定、免疫组织化学等技术进一步研究细胞外ATP通过P2X4受体诱发CIRI的分子机制,为缺血性脑中风治疗提供有益的理论依据

中文关键词： 脑缺血再灌注损伤；细胞外ATP；P2X4；受体；海马

英文摘要： The ischemic cerebral paralysis is the most common disease, which threatens human health severely.The important treatment is the restoration of local blood supply as soon as possible. But the neurons will further be injured in the ischemic region after restroration of blood supply, which is designated as cerebral ischemia reperfusion injury(CIRI). The mechanisms of CIRI are very complicated. The previous data showed that accumulation of extracellular toxins such as excitable amino acid, ATP, neuronal calcium overloading and radical generation etc are involved in the processes of CIRI. The previous data demonstrated that CIRI are often observed in CA1 region of hippocampus, cerebral cortex and striatum. Our preliminary data showed that the pyramidal neurons being apoptosis expressed high level of P2X4 receptor in the hippocampal CA1 region after global ischemia and reperfusion; the blockage of P2X4 receptor by 5BDBD(a selective antagonist for P2X4 ) could partly inhibited the apoptosis in this region; some of the neurons over-expressing P2X4 receptor via virus vectors in the CA1 region are also observed to be apoptosis. Those preliminary data of ours clearly indicated that up-regulation of P2X4 receptor after global ischemia and reperfusion is involved in the CIRI of hippocampal CA1 region. This research project will further investigate the mechanisms of CIRI induced by extracelluar ATP via P2X4 receptor by the following techniques such as tissue microanalysis, western blot, calcium assay and immunohistochemistry etc. The data from this reseach project will be useful for the treatment of the ischemic cerebral paralysis.

英文关键词： brain ischemia reperfusion injury;extracellular ATP;P2X4 receptor;hippocampus