Inflammatory control of blood-brain barrier integrity and epileptogenesis after seizures

癫痫发作后血脑屏障完整性和癫痫发生的炎症控制

基本信息

批准号：
9914359
负责人：
RAYMOND J DINGLEDINE
金额：
$ 42.22万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-15 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9914359
关键词：
Ablation Acute Address Albumins Alzheimer&apos s Disease Amygdaloid structure Amyotrophic Lateral Sclerosis Animal Model Anxiety Appearance Astrocytes Autoimmune Process Behavioral Assay Blood Blood - brain barrier anatomy Blood Vessels Brain Brain Injuries Capillary Endothelial Cell Cells Chronic Cognitive deficits Conditioned Culture Media Cortical Dysplasia Cyclic AMP-Dependent Protein Kinases Development Dinoprostone Disease Down-Regulation Electroencephalography Elements Encephalitis Endothelium Epilepsy Epileptogenesis Event Exposure to Flow Cytometry Fluorescein-5-isothiocyanate Genes Genetic Gliosis In Vitro Infection Infiltration Inflammation Inflammation Mediators Inflammatory Interleukin-1 Interleukin-6 Knockout Mice Life Maintenance Measures Mediating Mediator of activation protein Messenger RNA Microglia Modeling Modification Molecular Molecular Target Morbidity - disease rate Multiple Sclerosis Mus Neurodegenerative Disorders Neurological emergencies Neurons Neutrophilic Infiltrate PTGS2 gene Pathway interactions Pharmacology Pilocarpine Play Population Principal Investigator Production Prosencephalon Prostaglandin Receptor Proteins Quantitative Reverse Transcriptase PCR Reaction Refractory Rodent Role Seizures Series Serum Albumin Signal Pathway Status Epilepticus Stroke TNF gene Testing Up-Regulation Vascular Endothelial Cell Western Blotting Work aging brain arrestin 1 arrestin 2 cell type chemokine comorbidity conditional knockout cyclooxygenase 2 cytokine design in vitro Model in vivo interest kainate malformation man monocyte mouse model neuroinflammation neutralizing antibody novel object recognition prevent programs receptor recruit small molecule tool

项目摘要

Project Summary Accumulating evidence in animal models highlights that inflammation ensuing in the brain during status epilepticus (SE) may play a determinant role in ongoing seizures and their long-term detrimental consequences, independent of an infection or auto-immune cause. Studies in a multitude of animal models demonstrate that SE causes a rapid and intense inflammatory cascade in the forebrain involving interactions among neurons, reactive astrocytes, activated microglia, vascular endothelial cells and, eventually, infiltrating neutrophils and monocytes from the blood. The pathophysiological interactions among the various inflammatory molecules, and the sequence of events leading to their induction, have not yet been dissected. The broad cytokine burst and gliosis following SE is blunted in mice that have genetic ablations of COX-2 restricted to those principal forebrain neurons in which COX-2 is normally induced by SE, pointing to a role for COX-2 pathways in SE-induced inflammation including breakdown of the blood-brain barrier (BBB), which is sufficient to produce epilepsy. Previous work showed that the EP2 receptor mediates much of the COX-2 effect. We hypothesize that SE-related morbidity is largely due to activation of microglial EP2 receptors, which modulates production of cytokines that degrade the BBB. Our specific aims are: 1. To test the hypothesis that activated microglia rather than inflammatory monocytes are responsible for EP2- regulated BBB breakdown after seizures; 2. To test the hypothesis that IL- -secreted mediator and Epac the major EP2 signaling pathway that degrades the integrity of the blood-brain barrier after SE; 3. To determine whether EP2 activation plays a dominant role in the development of epilepsy or its comorbidities after SE. To address these aims we employ in vitro culture models of the BBB and in vivo SE models with cell- specific conditional knockouts of EP2. Immunohistochemical, western blot, FACS, qRT-PCR, EEG and behavioral assays are performed.

项目概要动物模型中不断积累的证据表明，癫痫持续状态期间大脑会发生炎症（SE）可能在持续癫痫发作及其长期有害后果中发挥决定性作用，独立感染或自身免疫原因。对多种动物模型的研究表明，SE 会导致快速前脑中强烈的炎症级联反应涉及神经元、反应性星形胶质细胞之间的相互作用，激活小胶质细胞、血管内皮细胞，并最终浸润中性粒细胞和单核细胞血。各种炎症分子之间的病理生理学相互作用以及事件的顺序导致他们的感应，尚未被解剖。 SE 后广泛的细胞因子爆发和神经胶质细胞增生被削弱在 COX-2 基因消融仅限于 COX-2 存在的主要前脑神经元的小鼠中通常由 SE 诱导，表明 COX-2 通路在 SE 诱导的炎症中发挥作用，包括血脑屏障（BBB）足以引起癫痫。先前的工作表明EP2受体介导大部分 COX-2 效应。我们假设 SE 相关的发病率很大程度上是由于激活小胶质细胞 EP2 受体，调节降解 BBB 的细胞因子的产生。我们的具体目标是：1. 为了检验激活的小胶质细胞而不是炎症单核细胞负责 EP2- 的假设癫痫发作后调节血脑屏障崩溃； 2. 检验IL- 分泌的假设介质和 Epac 是主要的 EP2 信号通路，在 SE 后会降低血脑屏障的完整性； 3. 确定EP2激活是否在癫痫或其合并症的发生中起主导作用在SE之后。为了实现这些目标，我们采用 BBB 的体外培养模型和具有细胞的体内 SE 模型 EP2 的特定条件敲除。免疫组织化学、蛋白质印迹、FACS、qRT-PCR、脑电图和行为学进行测定。