MECHANISMS OF ACTION OF IL 1 IN ISCHEMIC BRAIN

描述：(改编自申请者摘要)：本次活动的总体目标 研究是为了确定IL-1在大脑中的作用机制 缺血或再灌流后的损伤。脑损伤相关参数的研究 监测指标包括脑水肿、脑梗塞体积、脑血流量、 血脑屏障通透性和病理变化的研究 免疫组织化学和酶分析。炎症的迹象是 评估包括激活小胶质细胞和招募 中性粒细胞；此外，相关细胞因子水平和黏附 将对分子进行监测。最后，腺病毒-IL-1ra构建用于 用于在脑内过表达IL-1ra以抑制IL-1介导的效应 试图建立IL-1的产生和 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 将包括1)IL-1在体内的时间进程和位置的测定 2)IL-1释放与脑缺血的关系 和其他细胞因子(如IL-2、IL-6、IL-8、肿瘤坏死因子α)，3)IL-1的作用 在诱导细胞黏附分子(如ICAM-1和CD62E)和 循环炎症细胞(即髓过氧化物酶+)的浸润 中性粒细胞)，以及4)IL-1在聚集和激活中的作用 小胶质细胞。 具体目标2：确定IL-1是否对脑损伤起作用 暂时性大脑中动脉闭塞(TMCAO)。进行的研究 满足这一目标与目标1中提出的目标基本相同，但 将进行临时性MCAO，而不是永久性MCAO。因此， 在再灌流期间，要进行的测定包括：1) 小鼠大脑中动脉阻塞后IL-1β表达的时程变化 2)IL-1ra是否发生一过性过表达 腺病毒载体携带IL-1ra基因的脑室给药 减少再灌注性脑损伤；3)IL-1在脑缺血再灌流中的作用 细胞黏附分子和循环炎症细胞的浸润。

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.