Respiratory Endothelial Injury by Xanthine Oxide

黄嘌呤氧化物引起的呼吸内皮损伤

• 批准号: 6621351
• 负责人: JOHN E REPINE
• 金额: $ 38万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-10 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6621351
• 关键词: CD95 molecule; adult respiratory distress syndrome; aldehyde /ketone oxidoreductase; apoptosis; enzyme activity; enzyme mechanism; free radical oxygen; gene expression; interleukin 1; laboratory rat; lung injury; molecular pathology; oxidative stress; respiratory epithelium; xanthine oxidase

DESCRIPTION (provided by applicant): Dysfunction of lung epithelial cells - a key component of the alveolar-capillary barrier - is central to the development of acute lung injury (ARDS). Cytokines, such as interleukin-1 (IL-1), oxidative stress and FasL are increased in lungs of ARDS patients but their relationship to each other and ARDS is unknown. The sources of oxidative stress in ARDS patients are also unknown but aldehyde oxidase (AOX) and xanthine oxidoreductase (XOR), which is increased in ARDS patients, are intracellular oxygen radical (O2*) generating enzymes whose regulation might be of benefit in ARDS. Our data shows that: 1. Leak and inflammation increased in lungs of rats given IL-1 intratracheally 5h before in vivo. 2. XOR expression, allopurinol-inhibitable 02* production, and apoptosis increased in epithelial cells in lungs of rats given IL-1 intratracheally 24h before in vivo. 3. Inhibition of lung XOR/AOX activity by tungsten feeding decreased epithelial cell apoptosis in lungs of rats given IL-1 intratracheally 24h before in vivo. 4. XOR/AOX expression and allopurinol-inhibitable 02* production, but not apoptosis, increased in lung epithelial cells treated with IL-1 24h before in vitro. 5. IL-1 and 02* increased lung epithelial cell Fas expression in vitro. 6. Lung lavage from rats given IL-1 intratracheally 24h before contained FasL and caused apoptosis of lung epithelial cells in vitro that had increased Fas levels following IL-1 treatment 24h before in vitro. 7. XOR and AOX gene expression was increased in lung epithelial cells treated with IL-1/IL-6 in vitro. Our specific hypothesis is that increased IL-1 increases XOR and/or AOX activity in lung epithelial cells increasing lung epithelial cell O2* production and Fas expression. Concomitant IL-1 dependent increases in lung inflammation increase lung FasL levels and produce epithelial cell apoptosis which contributes to lung injury CARDS). Our specific aims are to determine the mechanisms responsible for IL-1 induced lung epithelial cell XOR and/or AOX expression, 02* production and epithelial cell apoptosis in vivo (Aim 1) and in vitro (Aim 2) and to determine the effect of IL-1 on the regulation of XOR and/or AOX gene expression in lung epithelial cells in vitro (Aim 3). The significance of this approach will be to determine basic physiologic, cellular and molecular aspects regarding XOR and AOX, to gain insight into whether XOR and/or AOX contribute to ARDS, and to evaluate whether inhibiting XOR and/or AOX holds any potential for treating or preventing events that contribute to ARDS.

描述（由申请方提供）：肺上皮细胞功能障碍- a 肺泡毛细血管屏障的关键组成部分-是发展的核心 急性肺损伤（ARDS）细胞因子，如白细胞介素-1（IL-1），氧化 ARDS患者肺内应激和FasL水平升高， 彼此之间，彼此之间，都是未知数。急性呼吸窘迫综合征的氧化应激来源 患者也未知，但醛氧化酶（AOX）和黄嘌呤 氧化还原酶（XOR），这是增加在ARDS患者，是细胞内 氧自由基（O2*）生成酶，其调节可能有益于 ARDS。 我们的数据显示：1.在给予的大鼠中， IL-1在体内实验前5 h，2.异或表达式， 在上皮细胞中，别嘌呤醇可降解的02* 产生和细胞凋亡增加。 在体内给药前24小时，大鼠肺内注射IL-1。3. 喂钨减少上皮细胞对肺XOR/AOX活性的抑制 IL-1在体内给药前24 h，肺组织细胞凋亡明显增加。 4. XOR/AOX表达和别嘌呤醇可降解的02* 产生，但不是 IL-1处理24 h后，肺上皮细胞凋亡增加， 体外5. IL-1和02* 在体外增加肺上皮细胞Fas表达。 6. IL-1治疗前24小时大鼠肺灌洗液中含有FasL 并在体外引起肺上皮细胞凋亡， IL-1处理后24小时的水平。7. XOR和AOX基因 在用IL-1/IL-6处理的肺上皮细胞中， 体外 我们的特定假设是，增加IL-1增加XOR和/或AOX 肺上皮细胞活性增加肺上皮细胞O2* 生产和Fas表达。伴随的肺中IL-1依赖性增加 炎症增加肺FasL水平并引起上皮细胞凋亡 这会导致肺损伤。 我们的具体目标是确定负责IL-1诱导 肺上皮细胞XOR和/或AOX表达、02* 产生和上皮细胞 在体内（目的1）和体外（目的2）细胞凋亡，并确定影响 IL-1对肺上皮细胞XOR和/或AOX基因表达的调节 体外细胞培养（Aim 3）。 这种方法的意义在于确定基本的生理， 关于XOR和AOX的细胞和分子方面，以深入了解 XOR和/或AOX是否有助于ARDS，并评估是否抑制 XOR和/或AOX具有治疗或预防以下事件的任何潜力， 导致ARDS。