Cytosolic Phospholipase A2 Alpha in Stroke Injury

胞浆磷脂酶 A2 Alpha 在中风损伤中的作用

• 批准号: 7149888
• 负责人: ADAM SAPIRSTEIN
• 金额: $ 35.88万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7149888
• 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; HTATIP gene; 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; PLA2G4A gene; PTGS2 gene; Partner in relationship; Pathway interactions; Perfusion; Permeability; Pharmaceutical Preparations; Phospholipase A2; Physiological reperfusion; Platelet Activating Factor; 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; cyclooxygenase 1; cyclooxygenase 2; dentate gyrus; deprivation; disability; doxorubicin/fluorouracil/melphalan protocol; gene induction; human PLA2G2A protein; human PLA2G4A protein; improved; in vitro Assay; inhibitor/antagonist; insight; iodoantipyrine; 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; size; stroke therapy; tert-Butylhydroperoxide; triphenyltetrazolium; vector

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A2 α (cPLA2a)在脑卒中后损伤演变中的作用。cPLA2a催化花生四烯酸从细胞膜向细胞内释放。脑缺血-再灌注激活磷脂酶A2，导致花生四烯酸及其类二十烷代谢产物水平升高。我们发现cpla2a缺陷小鼠在局灶性脑缺血/再灌注后神经元损伤明显减轻。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脑卒中损伤；(b) cPLA2a和KIDS- cPLA2a水平在脑缺血/再灌注后均升高，(c) cPLA2a和KIDS- cPLA2a之间的平衡影响缺血/再灌注后海马细胞的命运。目的1将测试cPLA2a敲除或接受特定cPLA2a抑制剂治疗的小鼠，与短暂性大脑中动脉闭塞后的对照组相比，是否会减少梗死面积、改变脑灌注和基因诱导。结果还将确定cPLA2a抑制的治疗时间窗。目的2将通过我们实验室开发的敏感分子、免疫组织化学和生化分析，将脑诱导cPLA2at类二十烷合成与缺血后炎症标志物联系起来。灵敏质谱法将识别cpla2a依赖性脂质介质的缺血/再灌注损伤。目的3将测试cPLA2a和KIDS- cPLA2a是否在小鼠短暂性全脑缺血后的海马中被诱导，并测量小鼠的分子和行为结果。在培养的海马神经元和器官型切片培养中，将使用我们实验室创建的慢病毒载体进行进一步检查。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酶的药物是否可以用于减少中风后的损伤，以及KIDS- cPLA2a蛋白是否有潜力成为一种新的脑特异性中风治疗方法。