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

• 批准号: 9151508
• 负责人: MARY C MCKENNA
• 金额: $ 25.57万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9151508
• 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; Creatine; Data; Development; Diagnosis; Energy Metabolism; Exhibits; Failure; Female; Glucose; Glutamates; Glutamine; Hippocampus (Brain); Hour; Image; Infant; Inflammation; Injury; Inositol; Lead; Learning; Lecithin; Left; 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; Neonatal; 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; Taurine; Techniques; Testing; Therapeutic; Time; 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; myelination; natural hypothermia; neonatal hypoxic-ischemic brain injury; neurotransmitter release; novel; perinatal injury; postnatal; protective effect; pup; response; synaptogenesis; white matter

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）导致皮层、皮层下和海马区的快速能量衰竭 这是由于缺乏氧气和葡萄糖输送到大脑。这之后是一个短暂的归一化， 再灌注和随后的二次能量衰竭（H/I后约2-6小时），最终导致脑损伤，以及 神经发育不良。在H/I之前发生的炎症可加重损伤。急性改变 新生儿H/I后的代谢和长期代谢失调使大脑脆弱， 支持正常发展所必需的进程。我们将使用围产期H/I的良好表征模型， 在有和没有先前炎症的情况下，在足月（出生后（PN）第10天大鼠幼仔）。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。我们将联合收割机高场1H-NMR和13 C-NMR光谱的能量 基质辅助激光解吸/电离-质谱法用于代谢和神经递质合成 在小脑的特定层中的代谢物、脂质、蛋白质的MALDI-MSI测定。我们 这项研究将是第一次确定通过星形胶质细胞和神经元特异性途径的代谢改变。 H/I后小脑谷氨酸和GABA的合成。分子和代谢的改变和长期- 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，导致小脑功能受损， 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的联合作用加剧了小脑损伤和失调， 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PN 10，有和没有预先炎症对雄性和雌性大鼠幼仔小脑能量代谢的影响。 2.确定浦肯野神经递质、代谢物、脂质和蛋白质的改变，分子和 H/I后雄性和雌性幼仔小脑的颗粒细胞层、深核和白色物质， 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， 以及这些化合物与低温联合对抗神经元和神经胶质代谢的变化， 浦肯野细胞、分子和颗粒细胞层、深层核和髓鞘中的特定分子变化。新颖、 关于神经元和星形胶质细胞损伤和改变的时间和靶点的临床相关信息 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 脂质和蛋白质在小脑层和深核将获得使用独特的组合1H- NMR和13 C-NMR结合MALDI-MSI和LC/MS-MS用于分析H/I后的变化， 无炎症、仅炎症和对神经保护治疗的反应。