MAP KINASE REGULATION OF MICROGLIAL ACTIVATION

描述（来自申请人的摘要）：小胶质细胞，主要的免疫细胞， 大脑的效应细胞，在调节大脑的功能中起着重要的作用。 CNS内的免疫微环境。它们会休眠直到 CNS的完整性受到损伤、感染或疾病过程的挑战 并且当慢性激活时，分泌大量炎症介质 包括细胞因子和组织损伤自由基作为致病因子的一部分 这是多种中枢神经系统疾病的共同机制，包括中风、阿尔茨海默氏病、 疾病、艾滋病痴呆和多发性硬化症等脱髓鞘疾病。一 对小胶质细胞活化过程的机械理解是， 因此，对于设计治疗策略来抑制 神经炎症该项目测试了信号转导的假设， 丝裂原活化蛋白激酶（MAPK）成员介导的通路 家族在微球激活和诱导 炎症反应。原代培养的大鼠脑小胶质细胞可作为 实现以下目标的模式。 MAPK级联的活性和作用（即，细胞外 信号调节激酶或ERK、p38 MAPK和c-Jun N-末端激酶或JNK） 将在响应内毒素激活的小胶质细胞中进行研究， 受体（CD 40）连接。MAPKs的药理学抑制剂对 小胶质细胞抗原的表达（即，MHC 11类、B7和CD 40）， 细胞因子（即，TNF α、IL-1、IL-6）和诱导型一氧化氮合酶（iNOS） 将通过免疫化学和RT-PCR技术确定。 p38 MAPK和JNK在肿瘤细胞中可能的亚型特异性作用和下游靶点 诱导细胞因子和iNOS基因表达的特征将是短暂的 使用激酶沿着iNOS的分子突变体的转染研究， 细胞因子基因启动子构建体。 激酶抑制剂对活化相关的 小胶质细胞功能;即，少突胶质细胞和髓磷脂的靶向 将使用体外模型测试吞噬作用。

Description (From the Applicant's Abstract): Microglia, the principal immune effector cells of the brain, play an important role in the regulation of the immunologic microenvironment within the CNS. They lie dormant until the integrity of the CNS is challenged by injury, infection or disease processes and when chronically activated, secrete a number of inflammatory mediators including cytokines and tissue damaging free radicals as part of the pathogenic mechanism common to a variety of CNS disorders including stroke, Alzheimer's Disease, AIDS dementia and demyelinating diseases such as multiple sclerosis. A mechanistic understanding of the process of microglial activation is, therefore, crucial for devising therapeutic strategies to suppress neuroinflammation. This project tests the hypothesis that signal transduction pathways mediated by members of the mitogen-activated protein kinase (MAPK) family play a key role in microgiial activation and the induction of inflammatory responses. Primary cultures of rat brain microglia wili be used as a model to accomplish the following objectives. The activities and the roles of MAPK cascades (i.e., extracellular signal-regulated kinase or ERK, p38 MAPK and c-Jun N-terminal kinase or JNK) will be investigated in microglia activated in response to endotoxin and receptor (CD40) ligation. The effects of pharmacological inhibitors of MAPKs on the expression of microglial antigens (i.e., MHC class 11, B7 and CD40), cytokines (i.e., TNFa, IL-1, IL-6) and inducible nitric oxide synthase (iNOS) will be determined by immunochemical and RT-PCR techniques. Possible isoforrn-specific roles and down-stream targets of p38 MAPK and JNK in inducing cytokine and iNOS gene expression will be characterized in transient transfection studies using molecular mutants of the kinases along with iNOS and cytokine gene promoter constructs. The suppressive effects of the kinase inhibitors on activation-associated microglial functions; i.e., targeting of oligodendrocytes and myelin phagocytosis will be tested using in vitro models.