脑缺血后胶质细胞铁代谢与炎症损伤机制研究及脑泰方干预作用

课题组前期研究证实铁超载参与了脑缺血神经元损伤，在此基础上，本项目提出脑缺血后神经胶质细胞铁代谢失衡与炎症反应密切相关；铁跨膜转运蛋白是脑缺血铁超载损伤的关键靶点，其表达受炎症反应调控；脑泰方可通过干预炎症反应调节铁跨膜转运蛋白表达抑制脑缺血后细胞内铁超载，发挥神经元保护作用。以MCAO模型大鼠为受试对象，运用免疫组化、PCR、WB等实验技术，检测分析缺血区小胶质细胞、星型胶质细胞的铁跨膜转运蛋白表达调控及神经肽（SP）、炎症因子（IL-1、IL-6、TNF-α）、“炎症--铁超载”信号通路BMP6/SMAD 和IL-6/STAT表达；体外实验，原代培养胶质细胞, 缺氧缺糖高铁处理后分组干预，检测铁跨膜转运蛋白及通路蛋白。阐明脑泰方对炎症诱导铁跨膜转运蛋白的表达干预铁转运的相关性以及调控信号转导机制的影响，为益气活血中药脑泰方防治缺血性脑损伤的临床应用和物质基础研究提供重要的实验依据。

Previous studies have demonstrated that iron overload is involved in cerebral ischemic injury. On the basis, the project put forward that iron metablism of glial cells is correlate to inflammation intimately ; The iron transmembrane protein is the key target of Cerebral ischemia injury caused by iron overload , and the expression of iron transmembrane protein is regulated by inflammatory reaction ; Naotaifang can intervene imflammation and protect the neurons by increasing the expression of iron transmembrane protein and inhibiting the overload of neuronal iron in cerebral ischemia. The MCAO model rats will be detected by experimental techniques such as Immuno histochemistry, PCR, WB and so on. These methods will be used to detect and alanysis the changes of intracellular iron , iron transmembrane proteins（TFR、DMT1、Fpn、FLVCR、BCRP）, Inflammatory factors （IL-6, IL-1β, TNF-α）and the expression of signal pathways related to “inflammation--iron overload mechanism”such as BMP6/SMAD and IL6/STAT . In the external experiment,we cultivate original glial cells with cultivate basement which is lack of oxygen and glucose but full of iron ion. Then we intervened them respectively and detected the iron transmembrane proteins and signal proteins by laser confocal microscope. The topic is in order to illustrate the correlation between iron transportion and the iron transmembrane protein expression , induced by inflammation and treated with Naotaifang . and my research concentrates on effect of Naotaifang on signal transduction mechanism of the iron transmembrane protein regulation . The research Provide an important experimental basis for clinical application and material basis about Naotaifang treating ischemic brain injury.