Glutamate Induced Molecular Events

描述(申请人提供)：我们令人兴奋的实验数据产生 在过去的几年中表明谷氨酸受体的新作用 介导的神经源性事件在炎症性伤害性反应中的关键作用 流程。脊髓背角给药的谷氨酸受体拮抗剂 消除大鼠炎症模型所致关节肿胀的一半。我们的 最近发现膝关节注射谷氨酸受体激动剂增加 关节血流量、血浆/蛋白质外渗、关节肿胀和神经 激活暗示外周谷氨酸的参与。这可能解释了 动物模型和临床患者炎症关节中谷氨酸的增加 有活动期的关节炎。在临床样本中，谷氨酸在患有 活动期关节炎，与患者其他关节或 在血浆中。这里提供的体外模型的初步数据表明 膝关节中细小的伤害性传入纤维释放谷氨酸 可通过刺激肿瘤直接启动炎症级联反应 加入特定谷氨酸激动剂后的肿瘤坏死因子(TNF_α)释放 滑膜细胞培养可增加培养物中检测到的肿瘤坏死因子-α水平 上清液。其次，观察到NMDA谷氨酸受体亚单位 在体外和体内模型中，NR1都可以转位到细胞核 提示可能参与了一种新的信号转导事件。把这些放在一起 这些发现导致了研究的假说，提出了谷氨酸 受体介导的分子事件在血管紧张素转换酶的启动中起关键作用 炎症性的级联反应。为这些研究设计的具体目标包括 具体目的：1.测定NMDA谷氨酸受体亚单位 NR1可以转位到细胞核。具体目标2.识别关键信号 调控NMDA谷氨酸受体亚单位核转位的途径 NR1。具体目的3.确定核NRI易位是否为 与谷氨酸诱导滑膜细胞释放肿瘤坏死因子cc有关。这些 研究将寻求可能导致新的翻译方法的机制 通过药物干预消除炎症状态 针对可诱导的谷氨酸受体介导的信号转导通路 因为使用谷氨酸受体拮抗剂不是临床选择。这些 研究将利用解剖学、生化和分子方法来检查 大鼠关节炎模型和人类克隆性滑膜培养。自案例 文献研究报告在某些病例中关节完整性恢复 中风后，这些研究试图更好地理解 在神经源性和免疫过程中希望逆转关节炎症 对病人来说是可能的。

DESCRIPTION (provided by applicant): Our exciting experimental data generated over the last several years indicate the novel role that glutamate receptor mediated neurogenic events play as a key factor in inflammatory nociceptive processes. Glutamate receptor antagonists given in the spinal cord dorsal horn eliminate half of the joint swelling induced in rat inflammation models. Our recent finding that knee injections of glutamate receptor agonists increase joint blood flow, plasma/protein extravasation, joint swelling, and nerve activation intimates peripheral glutamate involvement. This may account for glutamate increases in inflamed joints in animal models and in clinic patients with active arthritis. In clinical samples, glutamate is high in joints with active arthritis and is independent of levels in the patients' other joints or in the plasma. Preliminary data presented here for an in vitro model suggest that glutamate release by small, nociceptive afferent fibers in the knee joint can directly initiate inflammatory cascades through stimulation of tumor necrosis factor (TNF_alpha) release since specific glutamate agonists added to synovial cell cultures increase TNF_alpha levels detected in the culture supernatant. Secondly, the observation that the NMDA glutamate receptor subunit NR1 can translocate to the nucleus in both the in vitro and in vivo models suggests that a novel signal transduction event may be involved. Together these findings led to the hypothesis for the studies proposed that glutamate receptor_mediated molecular events play a critical role in initiation of inflammatory cascades. Specific Aims designed for these studies include the following: Specific Aim 1. To determine if the NMDA glutamate receptor subunit NR1 can translocate to the nucleus. Specific Aim 2. To identify key signaling pathways regulating nuclear translocation of NMDA glutamate receptor subunit NR1. Specific Aim 3. To determine if the nuclear NR I translocation is associated with glutamate_induced TNF_cc release from synoviocytes. These studies will seek mechanisms that may lead to novel translational approaches to abrogation of the inflammatory state through pharmacological interventions directed at inducible, glutamate receptor_mediated signal transduction pathways since use of glutamate receptor antagonists is not a clinical option. These studies will utilize anatomical, biochemical, and molecular methods to examine models of arthritis in rats and human clonal, synovial cultures. Since case studies in the literature report recovery of joint integrity in some cases after stroke, these studies seek a better understanding of the interactions of neurogenic and immune processes in the hope that reversal of joint inflammation for patients is possible.