MECHANISMS OF ACTION OF IL 1 IN ISCHEMIC BRAIN

描述：（改编自申请人的摘要）：本发明的总体目标是： 研究的目的是确定IL-1在脑中的作用机制 缺血或再灌注损伤。 脑损伤的参数 包括水肿、脑梗死体积、脑血流量， 血脑屏障渗透性，以及病理学变化， 免疫组织化学和酶测定。 炎症的迹象是 评估包括小胶质细胞的激活和 中性粒细胞;此外，相关细胞因子和粘附水平 分子是要监测的。 最后，腺病毒-IL-1 ra构建体将 用于在脑中过表达IL-1 ra以抑制IL-1介导的作用 为了试图建立IL-1产生与 刺激可能导致组织损伤的活动 在这些脑损伤模型中观察到。 因此，该提案的具体目标是：具体目标1：确定 IL-1在永久性中期脑损伤中的作用机制 脑动脉闭塞（pMCAO）。 为实现这一目标而进行的研究 包括确定1）IL-1在细胞中的时间过程和位置， 缺血小鼠脑组织中IL-1释放的关系 和其它细胞因子（例如，IL-2、IL-6、IL-8、TNF α），3）IL-1的作用 在诱导细胞粘附分子（例如，ICAM-1和CD 62 E）和 循环炎性细胞的浸润（即，髓过氧化物酶+ 中性粒细胞），和4）IL-1在积聚和激活中的作用， 小胶质细胞 具体目标2：确定IL-1是否有助于脑损伤， 暂时性大脑中动脉闭塞（tMCAO）。 进行的研究， 满足这一目标的基本上与目标1相同，除了 将执行临时而非永久MCAO。 因此，我们认为， 在再灌注期间，要进行的测定包括：1） tMCAO后小鼠脑中IL-1 β表达的时程， 2)IL-1 ra的瞬时过表达是否产生以下结果： 腺病毒介导的IL-1 ra基因脑室内递送 减少再灌注脑损伤，3）IL-1在诱导 细胞粘附分子和循环炎性细胞浸润。

DESCRIPTION: (Adapted from Applicant's Abstract): The overall goal of this investigation is to determine the mechanisms of action of IL-1 in brain injury following ischemia or reperfusion. The parameters of brain injury to be monitored include edema, cerebral infarction volume, cerebral blood flow, blood brain barrier permeability, and changes in pathology using methods of immunohistochemistry and enzyme assays. Signs of inflammation to be assessed include the activation of microglial cells and recruitment of neutrophils; in addition, levels of relevant cytokines and adherence molecules are to be monitored. Finally, Adenovirus-IL-1ra constructs are to be used to over-express IL-1ra in the brain to inhibit IL-1-mediated effects in an attempt to establish relationships between IL-1 production and the stimulation of activities that may be responsible for the tissue damage observed in these models of brain injury. Thus, the Specific Aims of the proposal are: Specific Aim 1: Identify the mechanism by which IL-1 contributes to brain injury during permanent middle cerebral artery occlusion (pMCAO). Studies performed to satisfy this aim are to include determinations of 1) the time course and location of IL-1 in the ischemic mouse brain, 2) the relationship between the release of IL-1 and other cytokines (e.g., IL-2, IL-6, IL-8, TNFalpha), 3) the role of IL-1 in induction of cell adhesion molecules (e.g., ICAM-1 and CD62E) and infiltration of circulating inflammatory cells (i.e., myeloperoxidase+ neutrophils), and 4) the role of IL-1 in the accumulation and activation of microglial cells. Specific Aim 2: Determine whether IL-1 contributes to brain injury during temporary middle cerebral artery occlusion (tMCAO). Studies performed to satisfy this aim are virtually the same as those presented for Aim 1 except the temporary instead of permanent MCAO is to be performed. Therefore, during reperfusion, determinations to be performed are to include 1) the time course for the expression of IL-1beta in mouse brain following tMCAO, 2) whether transient over-expression of IL-1ra produced following intraventricular delivery of the IL-1ra gene carried by adenovirus vectors decreases reperfusion brain injury, and 3) the role of IL-1 in induction of cell adhesion molecules and infiltration of circulating inflammatory cells.