12-Lipoxygenase and Ischemic Brain Cell Death

12-脂氧合酶和缺血性脑细胞死亡

基本信息

批准号：
7404413
负责人：
KLAUS VAN LEYEN
金额：
$ 33.55万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-04-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7404413
关键词：
12-HETE 12-HPETE Address Arachidonate 12-Lipoxygenase Attention Behavioral Blood - brain barrier anatomy Brain Brain Edema Brain Injuries Brain region Caspase Cell Death Cell Fractionation Cell Line Cell Survival Cells Cerebral Ischemia Cessation of life Complement Contralateral Cultured Cells Data Enzymes Extravasation Family Functional disorder Generations Genetic Glutamates Hippocampus (Brain)Hypoxia Immunofluorescence Immunologic Immunohistochemistry In Situ Nick-End Labeling Infarction Injury Ischemia Ischemic Brain Injury Knock-out Knockout Mice Knowledge LOX gene Lipid Peroxides Lipids Lipoxygenase Measures Mediating Mediator of activation protein Membrane Membrane Lipids Mitochondria Modeling Mus Neurons Organelles Other Finding Outcome Oxidative Stress PTGS2 gene Phospholipase Physiological reperfusion Polyunsaturated Fatty Acids Prostaglandin-Endoperoxide Synthase Protein Isoforms Protein-Lysine 6-Oxidase Proteins Rattus Reactive Nitrogen Species Reactive Oxygen Species Reperfusion Therapy Research Personnel Resistance Sampling Side Specificity Staining method Stains Stroke Techniques Testing Time Tissue Sample Toxic effect Up-Regulation Western Blotting Wild Type Mouse baicalein brain cell cell cortex cyclooxygenase 1 cyclooxygenase 2 cytotoxicity improved in vivo inhibitor/antagonist member mouse model multicatalytic endopeptidase complex neuron loss novel novel therapeutics programs research study size therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Lipoxygenases and cyclooxygenases are among the oxidative enzymes that contribute to the generation of reactive oxygen species (ROS) after stroke. While the cyclooxygenases COX-1 and COX-2 have been and are continuing to be extensively studied, little data is available on the contribution of lipoxygenases to ischemic brain cell death. Our preliminary results have shown that levels of 12-LOX, the major lipoxygenase isoform in mouse and rat brain, are elevated in a mouse model of focal ischemia, predominantly in neuronal cells of the cortex. 12-LOX knockout mice show significantly reduced infarction sizes, a finding that seems to be mimicked by treatment of wild-type mice with a 12-LOX inhibitor. In both rat primary neurons and a murine hippocampal cell line, we have confirmed and extended the findings of others that 12-LOX mediates oxidative stress-induced cell death, and neurons prepared from the 12-LOX knockout mice show enhanced resistance to this form of oxidative stress. We thus propose the following Specific Aims: 1. To investigate 12-LOX upregulation and changes in activity in mouse brain after ischemia with various times of reoxyenation and to correlate increased 12-LOX levels with the extent of brain damage; 2. To study the mechanisms by which 12-LOX contributes to the death of cultured neuronal cells, and to elucidate the degradative machinery responsible for executing cell death downstream of 12-LOX action; 3. To determine the consequences of inactivating 12-LOX by either genetic or pharmacological means on brain damage after focal ischemia in the mouse. With the knowledge gained from these studies we seek to establish 12-LOX as a novel therapeutic target in the treatment of stroke.

描述（由申请人提供）：脂氧合酶和环加氧酶属于氧化酶，有助于中风后产生活性氧（ROS）。虽然环加氧酶 COX-1 和 COX-2 已经并将继续得到广泛研究，但关于脂加氧酶对缺血性脑细胞死亡的贡献的数据很少。我们的初步结果表明，在局部缺血的小鼠模型中，12-LOX（小鼠和大鼠大脑中的主要脂氧合酶亚型）的水平升高，主要是在皮质神经元细胞中。 12-LOX 基因敲除小鼠的梗塞面积显着减小，这一发现似乎与用 12-LOX 抑制剂治疗野生型小鼠类似。在大鼠原代神经元和鼠海马细胞系中，我们已经证实并扩展了其他人的发现，即 12-LOX 介导氧化应激诱导的细胞死亡，并且从 12-LOX 敲除小鼠制备的神经元显示出对这种形式的增强的抵抗力。氧化应激。因此，我们提出以下具体目标： 1. 研究缺血后不同时间再氧合的小鼠大脑中 12-LOX 的上调和活性变化，并将 12-LOX 水平升高与脑损伤程度相关联； 2. 研究12-LOX导致培养神经元细胞死亡的机制，并阐明12-LOX作用下游执行细胞死亡的降解机制； 3. 确定通过遗传或药理学手段灭活 12-LOX 对小鼠局灶性缺血后脑损伤的影响。凭借从这些研究中获得的知识，我们寻求将 12-LOX 确立为治疗中风的新治疗靶点。