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酶胞质磷脂酶A2α（CPLA2A）在中风后损伤演变中的作用。 CPLA2A催化了从膜上蛛网膜酸的细胞内释放。脑缺血 - 再灌注会激活磷脂酶A2，导致蛛网膜酸及其类花生酸代谢物的水平增加。我们表明，局灶性脑缺血/再灌注后CPLA2A缺陷小鼠的神经元损伤明显减少。现在，我们已经确定了CPLA2A的Kids-cpla2a，这是一种新型的CPLA2A，该形式是在牙齿回和保护神经元中特异性诱导的。我们假设（a）CPLA2A活性通过扩增炎症来增强中风损伤；（b）脑缺血/再灌注后CPLA2A和CPLA2A的水平增加，（c）缺血/再灌注后，CPLA2A和CPLA2A之间的平衡会影响海马的细胞命运。 AIM 1将测试与瞬时大脑中动脉闭塞后对照组相比，与对照组相比，与对照组相比，与特定CPLA2A抑制剂治疗的CPLA2A敲除或接受特定CPLA2A抑制剂的小鼠是否降低了梗塞大小，大脑灌注和基因诱导。结果还将定义CPLA2A抑制的治疗时间窗口。 AIM 2将使用敏感的分子，免疫组织化学和生化化学测定法中开发的敏感分子，免疫组织化学分析，将炎症后CPLA2AT eicosanoid合成的大脑诱导和炎症标记相关联。敏感的质谱法将识别缺血/再灌注损伤的CPLA2A依赖性脂质介质。 AIM 3将测试在短暂全局缺血后，海马中诱导CPLA2A和KIDS-CPLA2A，并测量小鼠的分子和行为结果。将使用在我们实验室中创建的慢病毒载体进行培养和器官切片培养中海马神经元的进一步检查。中风损伤是一个主要的公共卫生问题，因为治疗的有效性有限。这些研究的结果将表明，针对CPLA2A酶的药物是否可以用于减少中风后的损伤，以及新蛋白（KIDS-CPLA2A）是否有潜力作为中风的新型脑特异性疗法。