Project II- Impact of Hypoxia-Ischemia and/or inflammation on Metabolism in Cerebellum

项目 II - 缺氧缺血和/或炎症对小脑代谢的影响

基本信息

批准号：
9979922
负责人：
MARY C MCKENNA
金额：
$ 25.65万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-17 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9979922
关键词：
ATP Synthesis Pathway Acute Address Astrocytes Attenuated Brain Brain Hypoxia-Ischemia Brain Injuries Brain region Cardiolipins Ceftriaxone Cell Death Cell Nucleus Cell Respiration Cell physiology Cerebellar Nuclei Cerebellum Cerebral Palsy Cerebral cortex Childhood Choline Citric Acid Cycle Clinical Creatine Data Development Diagnosis Energy Metabolism Exhibits Failure Female Glucose Glutamates Glutamine Hippocampus (Brain)Hour Image Impairment Infant Inflammation Injury Inositol Lead Learning Lecithin Lipids Lysophospholipids Mass Spectrum Analysis Membrane Membrane Lipids Membrane Microdomains Metabolic Metabolic Pathway Metabolism Mitochondria Modeling Molecular Motor Motor Skills Myelin N-acetylaspartate NADP NMR Spectroscopy Neurites Neurons Neurotransmitters Nucleotides Outcome Oxidation-Reduction Oxidative Stress Oxygen Pathway interactions Pentosephosphate Pathway Perinatal Hypoxia Pharmacotherapy Phosphocreatine Phospholipids Process Production Protein Biosynthesis Proteins Purkinje Cells Pyruvate Carboxylase Rattus Reduced Glutathione Reperfusion Therapy Signal Transduction Specificity Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Sphingomyelins Structure Taurine Techniques Testing Therapeutic behavioral outcome brain cell brain metabolism clinically relevant functional outcomes gamma-Aminobutyric Acid granule cell improved outcome insight lipid biosynthesis liquid chromatography mass spectrometry male membrane synthesis myelination natural hypothermia neonatal hypoxic-ischemic brain injury neonatal period neurotransmitter release novel perinatal injury postnatal protective effect pup response synaptogenesis white matter

项目摘要

Neonatal hypoxia-ischemia (H/I) leads to rapid energy failure in cortical, subcortical and hippocampal regions due to lack of oxygen and glucose delivery to brain. This is followed by a transient normalization during reperfusion and a later secondary energy failure (~2-6 hours after H/I) that ultimately leads to brain injury, and poor neurodevelopmental outcome. Inflammation occurring prior to H/I can exacerbate injury. Acute alterations in metabolism and prolonged metabolic dysregulation after neonatal H/I leave the brain vulnerable and unable to support processes essential for normal development. We will use a well-characterized model of perinatal H/I at term (postnatal (PN) day 10 rat pup) with and without prior inflammation. The effects of H/I on cerebellum have not been carefully explored even though the rapidly developing cerebellum is particularly vulnerable to inflammation and perinatal injury. We will combine high field 1H-NMR and 13C-NMR spectroscopy of energy metabolism and neurotransmitter synthesis with matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) determination of metabolites, lipids, proteins in specific layers of the cerebellum. Our studies will be the first determination of alterations in metabolism via astrocyte and neuron specific pathways and synthesis of glutamate and GABA in cerebellum after H/I. Molecular and metabolic alterations and long- term functional outcomes will determine the therapeutic potential of hypothermia and drug therapy. We hypothesize that neonatal H/I leads to energy failure in the cerebellum that contributes to impaired function of brain cells including acute and long-term alterations in energy metabolism and neurotransmitter synthesis. We hypothesize that the combined effect of inflammation and H/I exacerbates cerebellar damage and dysregulated metabolism in male and female brain. Our Specific Aims test these hypotheses: 1. Determine the effect of H/I at PN10 with and without prior inflammation on energy metabolism in cerebellum of male and female rat pups. 2. Determine the alterations in neurotransmitters, metabolites, lipids and proteins in Purkinje, molecular and granule cell layers, deep nuclei and white matter of the cerebellum of male and female pups after H/I with and without prior inflammation. 3. Determine the protective effect of hypothermia, choline and ceftriaxone alone, and these compounds in combination with hypothermia against changes in neuronal and glial metabolism and specific molecular changes in the Purkinje, molecular and granule cell layers, deep nuclei and myelin. Novel, clinically relevant information about the timing and targets of injury and alterations in neuron and astrocyte specific metabolic pathways in brain and alterations of metabolites, nucleotides, membrane and signaling lipids, and proteins in cerebellar layers and deep nuclei will be obtained using the unique combination of 1H- NMR and 13C-NMR in conjunction with MALDI-MSI and LC/MS-MS for profiling the changes after H/I with and without inflammation, inflammation alone and response to neuroprotective therapy.

新生儿缺氧缺血（H/I）导致皮质、皮质下和海马区域快速能量衰竭由于缺乏氧气和葡萄糖输送到大脑。接下来是瞬态标准化期间再灌注和随后的继发性能量衰竭（H/I 后约 2-6 小时）最终导致脑损伤，以及神经发育结果不佳。 H/I 之前发生的炎症会加剧损伤。急性改变新生儿 H/I 后的新陈代谢和长期代谢失调使大脑变得脆弱且无法支持正常开发所必需的流程。我们将使用一个特征良好的围产期 H/I 模型足月（出生后 (PN) 第 10 天的幼鼠）有或没有先前的炎症。 H/I 对小脑的影响尽管快速发育的小脑特别容易受到炎症和围产期损伤。我们将结合高场 1H-NMR 和 13C-NMR 能量谱使用基质辅助激光解吸/电离质谱法进行代谢和神经递质合成成像（MALDI-MSI）测定小脑特定层的代谢物、脂质、蛋白质。我们的研究将首次确定星形胶质细胞和神经元特异性途径代谢的改变 H/I 后小脑中谷氨酸和 GABA 的合成。分子和代谢的改变以及长期长期功能结果将决定低温和药物治疗的治疗潜力。我们假设新生儿 H/I 导致小脑能量衰竭，从而导致小脑功能受损脑细胞，包括能量代谢和神经递质合成的急性和长期改变。我们假设炎症和 H/I 的联合作用会加剧小脑损伤和失调男性和女性大脑的新陈代谢。我们的具体目标测试这些假设： 1. 确定 H/I 的影响在 PN10 时，无论有无先前炎症，雄性和雌性幼鼠小脑能量代谢的变化。 2. 确定浦肯野、分子和蛋白质中神经递质、代谢物、脂质和蛋白质的变化 H/I 后雄性和雌性幼崽小脑的颗粒细胞层、深部核团和白质之前没有炎症。 3. 确定单独使用低温、胆碱和头孢曲松的保护作用，这些化合物与低温相结合，可以对抗神经元和神经胶质代谢的变化，浦肯野、分子和颗粒细胞层、深部细胞核和髓磷脂的特定分子变化。小说，有关神经元和星形胶质细胞损伤和改变的时间和目标的临床相关信息大脑中的特定代谢途径以及代谢物、核苷酸、膜和信号传导的改变小脑层和深核中的脂质和蛋白质将使用 1H- 的独特组合获得 NMR 和 13C-NMR 与 MALDI-MSI 和 LC/MS-MS 结合使用和分析 H/I 后的变化无炎症、仅炎症且对神经保护治疗有反应。