Cytosolic Phospholipase A2 Alpha in Stroke Injury

胞浆磷脂酶 A2 Alpha 在中风损伤中的作用

基本信息

批准号：
7899946
负责人：
ADAM SAPIRSTEIN
金额：
$ 35.52万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-01 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7899946
关键词：
Abbreviations Achievement Acute Aminoisobutyric Acids Arachidonic Acids Area Autoradiography Behavioral Biochemical Biological Assay Blood - brain barrier anatomy Blood Platelets Brain Brain Injuries Cause of Death Cell Count Cells Cerebral Ischemia Cerebrovascular Circulation Cerebrum Chloride Ion Chlorides Clinical Treatment Cytosolic Phospholipase A2 Eicosanoids Enzyme-Linked Immunosorbent Assay Enzymes Equilibrium Evolution Genes Genetic Genotype Glucose Hippocampus (Brain)Histologic Immunohistochemistry Infarction Infiltration Inflammation Inflammatory Injury Ischemia Ischemic Stroke Kainic Acid Knock-out Knockout Mice Lasers Lentivirus Vector Leukocytes Link Lipids Mass Spectrum Analysis Measures Membrane Messenger RNA Methods Microglia Middle Cerebral Artery Occlusion Modeling Molecular Molecular Biology Mus Neuronal Injury Neurons Outcome Oxygen PLA2G2A gene PTGS2 gene Partner in relationship Pathway interactions Perfusion Permeability Pharmaceutical Preparations Phospholipase A2 Prostaglandin-Endoperoxide Synthase Prostaglandins Proteins Public Health Reperfusion Injury Reperfusion Therapy Research Personnel Role Serum Slice Solutions Stroke Subfamily lentivirinae System Techniques Testing Therapeutic Time Toxic effect Treatment Effectiveness behavior test butyrine dentate gyrus deprivation disability gene induction human PLA2G4A protein improved in vitro Assay inflammatory marker inhibitor/antagonist insight kainate lipid mediator middle cerebral artery mouse model neuron loss neuronal survival neutrophil novel oxygen toxicity progesterone 11-hemisuccinate-(2-iodohistamine)programs response salt balance stroke therapy tert-Butylhydroperoxide triphenyltetrazolium vector

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this project is to determine the roles of the enzyme cytosolic phospholipase A2 alpha (cPLA2a) in the evolution of injury following stroke. cPLA2a catalyzes the intracellular release of arachidonic acid from membranes. Cerebral ischemia-reperfusion activates phospholipase A2 resulting in increased levels of arachidonic acid and its eicosanoid metabolites. We showed that cPLA2a-deficient mice suffer significantly less neuronal injury after focal cerebral ischemia/reperfusion. We have now identified KIDS- cPLA2a, a novel form of cPLA2a, which is specifically induced in the dentate gyrus and protects neurons. We hypothesize (a) that cPLA2a activity enhances stroke injury by amplifying inflammation; (b) that levels of both cPLA2a and KIDS- cPLA2a are increased following cerebral ischemia/reperfusion, (c) that the balance between cPLA2a and KIDS- cPLA2a influences cell fate in the hippocampus following ischemia/reperfusion. Aim 1 will test if cPLA2a knockouts or mice treated with specific cPLA2a inhibitors have decreased infarct size, altered cerebral perfusion, and gene induction, as compared to controls after transient middle cerebral artery occlusion. Results will also define a therapeutic time window for cPLA2a inhibition. Aim 2 will correlate the cerebral induction of cPLA2at eicosanoid synthesis, and markers of inflammation following ischemia using sensitive molecular, immunohistochemical and biochemical assays developed in our labs. Sensitive mass spectrometry will identify cPLA2a-dependent lipid mediators of ischemia/reperfusion injury. Aim 3 will test if cPLA2a and KIDS- cPLA2a are induced in the hippocampus following transient global ischemia and measure molecular and behavioral outcomes in mice. Further examination in hippocampal neurons in culture and in organotypic slice culture will be performed using lentivirus vectors created in our lab. Stroke injury is a major public health problem because of the limited effectiveness of treatments. The results of these studies will show if drugs aimed against the enzyme cPLA2a can be used to decrease injury following stroke and if the new protein, KIDS- cPLA2a has potential as a new brain-specific therapy for stroke.

描述（由申请人提供）：该项目的长期目标是确定胞浆磷脂酶 A2 α (cPLA2a) 在中风后损伤演变中的作用。 cPLA2a 催化细胞内膜上花生四烯酸的释放。脑缺血再灌注会激活磷脂酶 A2，导致花生四烯酸及其类二十烷酸代谢物水平增加。我们发现 cPLA2a 缺陷小鼠在局灶性脑缺血/再灌注后遭受的神经元损伤明显减少。我们现在已经鉴定出 KIDS-cPLA2a，这是 cPLA2a 的一种新形式，它在齿状回中被特异性诱导并保护神经元。我们假设 (a) cPLA2a 活性通过放大炎症来增强中风损伤； (b) cPLA2a 和 KIDS-cPLA2a 的水平在脑缺血/再灌注后增加，(c) cPLA2a 和 KIDS-cPLA2a 之间的平衡影响缺血/再灌注后海马中的细胞命运。目标 1 将测试与短暂大脑中动脉闭塞后的对照组相比，cPLA2a 敲除小鼠或用特定 cPLA2a 抑制剂治疗的小鼠是否减少了梗塞面积、改变了脑灌注和基因诱导。结果还将定义 cPLA2a 抑制的治疗时间窗。目标 2 将使用我们实验室开发的敏感分子、免疫组织化学和生化检测将 cPLA2at 类二十烷酸合成的大脑诱导与缺血后的炎症标志物关联起来。灵敏的质谱分析将鉴定缺血/再灌注损伤的 cPLA2a 依赖性脂质介质。目标 3 将测试短暂性整体缺血后海马中是否会诱导 cPLA2a 和 KIDS-cPLA2a，并测量小鼠的分子和行为结果。将使用我们实验室创建的慢病毒载体对培养物和器官切片培养物中的海马神经元进行进一步检查。由于治疗效果有限，中风损伤是一个主要的公共卫生问题。这些研究的结果将表明，针对 cPLA2a 酶的药物是否可用于减少中风后的损伤，以及新蛋白质 KIDS-cPLA2a 是否有潜力成为中风的新的大脑特异性疗法。