MAP KINASE REGULATION OF MICROGLIAL ACTIVATION

MAP 激酶对小胶质细胞激活的调节

• 批准号: 6724927
• 负责人: NARAYAN R BHAT
• 金额: $ 21.45万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-15 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6724927
• 关键词: CD40 molecule; JUN kinase; MHC class II antigen; cell cell interaction; endotoxins; enzyme activity; enzyme induction /repression; enzyme inhibitors; immunocytochemistry; interleukin 1; interleukin 6; laboratory rat; microglia; mitogen activated protein kinase; mixed tissue /cell culture; myelin; neuroimmunomodulation; nitric oxide synthase; oligodendroglia; phagocytosis; polymerase chain reaction; surface antigens; tumor necrosis factor alpha

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.

描述（摘自申请人的摘要）：小胶质细胞，主要免疫 大脑的效应细胞在调节中起重要作用 中枢神经系统内的免疫学微环境。他们处于休眠状态直到 中枢神经系统的完整性受到伤害，感染或疾病过程的挑战 当长期激活时，分泌许多炎症介体 包括细胞因子和组织破坏自由基作为致病性的一部分 各种中枢神经系统疾病（包括中风），阿尔茨海默氏症的常见机制 疾病，艾滋病痴呆症和脱髓鞘疾病，例如多发性硬化症。一个 对小胶质激活过程的机械理解是， 因此，对于制定抑制治疗策略至关重要 神经炎症。该项目检验了信号转导的假设 有丝分裂原激活蛋白激酶（MAPK）介导的途径 家庭在小胶质激活和诱导中起关键作用 炎症反应。大鼠脑小胶质细胞的一级培养被用作 实现以下目标的模型。 MAPK级联的活动和角色（即细胞外 信号调节的激酶或ERK，p38 MAPK和C-JUN N末端激酶或JNK） 将在响应内毒素和 受体（CD40）连接。 MAPK药理抑制剂对 小胶质细胞抗原的表达（即MHC类11，B7和CD40）， 细胞因子（即TNFA，IL-1，IL-6）和可诱导的一氧化氮合酶（INOS） 将由免疫化学和RT-PCR技术确定。 p38 mapk和jnk的可能的异金特异性角色和下游目标 诱导细胞因子和iNOS基因表达将在瞬态中进行表征 转染研究使用激酶的分子突变体以及iNOS和 细胞因子基因启动子构建体。 激酶抑制剂对激活相关的抑制作用 小胶质函数；即，寡胶质细胞和髓鞘的靶向 吞噬作用将使用体外模型进行测试。