CRITICAL TARGETS IN HYPEROXIC MITOCHONDRIAL INJURY

高氧线粒体损伤的关键目标

• 批准号: 7716163
• 负责人: Carl W White
• 金额: $ 2.26万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7716163
• 关键词: Aconitate Hydratase; Adenine Nucleotides; Adult; Antioxidants; Binding; Biochemical; Biological Assay; Bronchopulmonary Dysplasia; Cell Respiration; Chronic lung disease; Complex; Computer Retrieval of Information on Scientific Projects Database; Continuous Infusion; Electron Microscopy; Environmental air flow; Enzymes; Exposure to; Funding; Glutamine; Glutathione; Glycolysis; Grant; Histopathology; Hyperoxia; Injury; Institution; Lung; Measures; Mitochondria; Modeling; Newborn Infant; Oxidants; Oxidative Stress; Oxygen; Papio; Premature Birth; Proteins; Rate; Rattus; Research; Research Personnel; Resources; Respiratory distress; S-Adenosylmethionine; Source; Stress; Sulfur Amino Acids; Superoxide Dismutase; Supplementation; TXN gene; Testing; Thioredoxin; Transcriptional Activation; United States National Institutes of Health; Up-Regulation; Voltage-Dependent Anion Channel; base; hexokinase; porin

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bronchopulmonary dysplasia (BPD) is a common, disabling and sometimes fatal chronic lung disease. It frequently accompanies premature birth and treatment of respiratory distress with artificial ventilation and high concentrations of inspired oxygen (hyperoxia). The biochemical basis of BPD is not well understood. In the baboon, exposure to hyperoxia is required treatment in both the ultra-premature (125 d) and premature (140) model of BPD. Hyperoxia damages mitochondria results in loss of aconitase activity, decreased mitochondrial and cell respiration, and loss of ATP. On that basis, this proposal's principal hypothesis is that adaptation to hyperoxic stress requires up-regulation of glycolytic, and/or glutaminolytic, enzymes. Because hexokinase rate- limits glycolysis in lung, it is hypothesized that expression of lung hexokinase(s) is up-regulated. Replacement of deficient mitochondrial anti-oxidants may alleviate early respiratory distress and resulting BPD and decrease up regulation of glycolytic enzymes. These hypotheses will be tested in these AIMS; 1) Determine differential expression of mRNA's encoding components of the mitochondrial porin complex-hexokinases (HKs), porins, and adenine nucleotide translocators (ANT), these same proteins, and relevant glycolytic and glutaminolytic enzymes; 2) Define early status of critical anti-oxidants (glutathione [GSH], thioredoxin [TRX], superoxide dismutases]), their precursors (S-adenosylmethionine), and oxidant target (aconitase) markers, and (3) Define the efficacy of early, continuous infusion of S-adenosylmethionine (AdoMet), a precursor of cellular and mitochondrial glutathione (GSH), on these markers and pulmonary histopathology of BPD. Hexokinase binding to, or release from, mitochondria will be quantitied using immunogold electron microscopy. Activities of HK, phosphofructokinase, and additional critical glycolytic and glutaminolytic enzymes also will be assayed. Lung GSH, TRX, and AdoMet, as well as circulating GSH, AdoMet, and sulfur amino acids will be measured early in the 125 d model. Together, these approaches will help define whether changes in hexokinase activity expression, known to occur in lungs of adult rats made oxygen-tolerant, also occur in the premature newborn baboon. In addition, they will indicate whether or not increased expression of pulmonary glutamine- utilizing enzymes also occurs during pulmonary oxidative stress and whether anti-oxidant stress and whether anti-oxidant supplementation modifies these adaptations.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 支气管肺发育不良（BPD）是一种常见，残疾，有时是致命的慢性肺部疾病。它经常伴随着人工通风和高浓度的氧气（高氧）的过早出生和治疗呼吸窘迫。 BPD的生化基础尚不清楚。在狒狒中，在BPD的超新生（125 d）和过早（140）模型中，需要接触高氧。高氧损害线粒体会导致刺刺酶活性的丧失，线粒体呼吸和细胞呼吸降低以及ATP的丧失。在此基础上，该提议的主要假设是对高氧化应激的适应需要上调糖酵解和/或谷氨酰胺分解酶。由于己糖酶速率 - 限制了肺中的糖酵解，因此假设肺己糖激酶的表达被上调。替换缺乏线粒体抗氧化剂可能会减轻早期呼吸窘迫，并导致BPD并减少糖酵解酶的调节。这些假设将在这些目标中进行检验。 1）确定mRNA的编码组件的差异表达，即线粒体孔蛋白复合 - 己糖苷（HKS），porins和腺嘌呤核苷酸易位剂（ANT），这些相同的蛋白质以及相关的糖酵解和谷氨酰胺分解酶； 2) Define early status of critical anti-oxidants (glutathione [GSH], thioredoxin [TRX], superoxide dismutases]), their precursors (S-adenosylmethionine), and oxidant target (aconitase) markers, and (3) Define the efficacy of early, continuous infusion of S-adenosylmethionine (AdoMet), a precursor of cellular and线粒体谷胱甘肽（GSH），在这些标记和BPD的肺部组织病理学上。己糖苷酶的结合或使用免疫元电子显微镜定量或从线粒体释放。还将分析HK，磷酸果糖激酶以及其他关键的糖酵解和谷氨酰胺分解酶的活性。肺GSH，TRX和ADOMEN以及循环GSH，ADOMET和硫氨基酸将在125 d模型的早期进行测量。这些方法共同有助于定义己糖激酶活性表达的变化是否发生在成年大鼠的肺中，使耐氧剂也发生在早产狒狒中。此外，它们将表明在肺部氧化应激期间还会发生肺谷氨酰胺的表达增加，以及抗氧化应激以及抗氧化剂补充是否会改变这些适应性。