MAP KINASE REGULATION OF MICROGLIAL ACTIVATION

描述(摘自申请者摘要)：小胶质细胞，主要免疫细胞 大脑的效应细胞，在调节 中枢神经系统内的免疫微环境。它们休眠着，直到 中枢神经系统的完整性受到损伤、感染或疾病过程的挑战 当长期激活时，会分泌一些炎症介质 包括细胞因子和组织破坏性自由基作为致病因素的一部分 多种中枢神经系统疾病的共同机制，包括中风、阿尔茨海默氏症 疾病、艾滋病、痴呆症和多发性硬化症等脱髓鞘疾病。一个 对小胶质细胞激活过程的机械理解是， 因此，制定治疗策略对抑制 神经炎。这个项目测试了信号转导的假设 丝裂原活化蛋白激酶(MAPK)成员的信号转导途径 家庭在微生物的激活和诱导中起着关键作用 炎症反应。大鼠脑小胶质细胞的原代培养 一个实现以下目标的模型。 MAPK级联(即细胞外)的活性和作用 信号调节激酶或ERK、p38MAPK和c-jun氨基末端激酶或JNK) 将在内毒素激活的小胶质细胞中进行研究 受体(CD40)结扎。MAPKs的药理抑制剂对血管内皮细胞生长的影响 小胶质细胞抗原(即MHC 11、B7和CD40)的表达； 细胞因子(TNFa、IL-1、IL-6)与诱导型一氧化氮合酶(INOS) 将通过免疫化学和RT-PCR技术进行检测。 P38 MAPK和JNK可能的异构体特异性作用和下游靶点 诱导细胞因子和诱导型一氧化氮合酶基因表达的特征是 利用激酶分子突变体以及iNOS和iNOS进行的转基因研究 细胞因子基因启动子构建。 激酶抑制剂对激活相关蛋白的抑制作用 小胶质细胞功能；即靶向少突胶质细胞和髓鞘 吞噬作用将使用体外模型进行测试。

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.