Microdialysis Studies of Seizure-Induced Oxidative Stress

癫痫发作引起的氧化应激的微透析研究

• 批准号: 8420439
• 负责人: CRAIG E LUNTE
• 金额: $ 31.03万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-16 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8420439
• 关键词: 8-hydroxy-2' -deoxyguanosine; 8-hydroxyguanosine; Acids; Affect; Amino Acid Neurotransmitters; Antioxidants; Arachidonic Acids; Biochemical Pathway; Biological Markers; Brain; Brain region; Butyric Acids; Catecholamines; Chemicals; Coenzyme Q10; Convulsions; Coupled; DNA; DNA Damage; Disulfides; Dose; Epilepsy; Esters; Event; Experimental Models; Fatty Acids; Generalized seizures; Glutamates; Glutathione; Goals; Indomethacin; Kindling (Neurology); Laboratories; Lead; Lipid Peroxidation; Lipids; Malondialdehyde; Measures; Membrane; Metabolism; Methodology; Methods; Microdialysis; Modeling; Monitor; Neurologic; Nitrogen; Oxidative Stress; Oxygen; Pentylenetetrazole; Phospholipase A2; Plasma; Population; Principal Investigator; Prostaglandins; Proteins; Reactive Nitrogen Species; Reactive Oxygen Species; Research; Role; Sampling; Seizures; Status Epilepticus; Sulfhydryl Compounds; Techniques; Time; Tissue Sample; Urine; Wistar Rats; analytical method; buthionine; cyclooxygenase 1; diethyl maleate; gamma-Aminobutyric Acid; in vivo; insight; oxidation; oxidative damage; programs; research study; response; sample collection

DESCRIPTION (provided by applicant): The purpose of this project is to investigate the role of epileptic seizures in oxidative stress. There is a wealth of research that demonstrates that epileptic seizures lead to the formation of reactive oxygen and nitrogen species (ROS and RNS), which cause oxidative damage to DNA, lipids, and proteins. The role of excitotoxic events during epileptic seizures and the subsequent formation of ROS and RNS leading to oxidative stress will be investigated. Microdialysis sampling in discrete brain regions along with simultaneous recording of electrocorticographic (ECoG) activity will be used to probe chemical and electrographic activity changes in the brain associated with seizures. This will provide the ability to correlate biomarker levels with seizure activity. Microdialysis sampling will be used to continuously monitor several biochemical pathways prior to, during, and after induction of seizures. Microdialysis experiments will provide both temporal and spatial information about oxidative stress caused by seizures and the brains response to them. This approach will provide insight into questions that have remained unclear using plasma and urine sampling and standard tissue sampling techniques. Methods previously developed in the PI's laboratory will be used to monitor formation of ROS and RNS, biomarkers for DNA damage and lipid peroxidation, and the neurotransmitter amino acids and catecholamines. A new analytical method will be developed to detect prostanoids resulting from arachidonic acid metabolism. Three experimental models of epilepsy will be used in Wistar rats. The first two models use 3-mercaptopropionic acid (3-MPA) to chemically induce seizures in a controlled manner. For one model the 3-MPA is systemically administered and in the other it is locally administered through the microdialysis probe. A chemical kindling model will then be used, where a sub-threshold dose (for triggering seizures) of pentylenetetrazol will be used to generate seizures of increasing intensity, culminating in a generalized seizure convulsion. In addition, we propose to investigate the role of various modulation agents on the release of known oxidative stress agents during epileptic seizures. Using microdialysis sampling coupled to the advanced analytical methodology, the neurological events leading from seizures to oxidative stress will be investigated. In particular, the temporal relationship between oxidative stress (as measured by ROS and RNS formation), and neuroexcitation (as measured by the GABA/glutamate ratio and the catecholamines) will be determined and correlated to the duration and intensity of seizure episodes.

描述（由申请人提供）：该项目的目的是研究癫痫发作在氧化应激中的作用。大量研究表明，癫痫发作会导致反应性氧和氮种（ROS和RNS）的形成，这会对DNA，脂质和蛋白质造成氧化损害。将研究兴奋性事件在癫痫发作中的作用以及随后导致氧化应激的ROS和RN的形成。离散大脑区域中的微透析采样以及电皮质学（ECOG）活性的同时记录将用于探测与癫痫发作相关的大脑中的化学和电视活性变化。这将提供将生物标志物水平与癫痫活性相关的能力。微透析抽样将用于 在癫痫发作之前，期间和之后，连续监测几种生化途径。微透析实验将提供有关癫痫发作以及大脑对其的氧化应激的时间和空间信息。这种方法将提供有关使用血浆和尿液采样以及标准组织采样技术一直不清楚的问题的洞察力。 先前在PI实验室开发的方法将用于监测ROS和RN的形成，用于DNA损伤和脂质过氧化的生物标志物以及神经递质氨基酸和儿茶酚胺。将开发一种新的分析方法来检测蛛网膜酸代谢引起的前列腺素。 Wistar大鼠将使用三个实验模型的癫痫模型。前两个模型使用3-甲基丙酸（3-MPA）以受控方式诱导癫痫发作。对于一种模型，3-MPA是系统地给药的，另一个模型是通过微透析探针局部给药的。然后，将使用化学点燃模型，其中将使用亚阈值剂量（用于触发癫痫发作）的甲状酸乙酸酯唑酚来产生增加的癫痫发作 强度，最终在广泛的癫痫发作中。此外，我们建议研究各种调节剂在癫痫发作期间释放已知氧化应激剂的作用。使用与晚期分析方法结合的微透析采样，将研究从癫痫发作到氧化应激的神经系统事件。特别是，将确定并与癫痫发作的持续时间和强度相关联，氧化应激（通过ROS和RNS形成）与神经兴奋（通过GABA/谷氨酸比和儿茶酚胺测量）之间的时间关系。