OXIDANTS AND ENDOTOXIN INDUCED ENDOTHELIAL INJURY

氧化剂和内毒素引起的内皮损伤

• 批准号: 6184078
• 负责人: BARBARA O MEYRICK
• 金额: $ 31.09万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-07-01 至 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6184078
• 关键词: cell death; cyclic GMP; endotoxins; enzyme activity; enzyme induction /repression; glutamine; nitric oxide; nitric oxide synthase; northern blottings; oxidative stress; peroxynitrites; prostaglandin endoperoxide synthase; prostaglandins; protein kinase C; tissue /cell culture; vascular endothelium; western blottings

DESCRIPTION (Adapted from the applicant's abstract): The applicant will further pursue the observation that intracellular generation of reactive oxygen species (ROS) contributes to endotoxin-stimulated prostanoid synthesis and release from pulmonary endothelium. Two closely associated hypotheses are proposed based on preliminary data demonstrating that endotoxin causes intracellular generation of ROS, and that the antioxidant, DMSO, as well as extracellularly generated nitric oxide (NO) and peroxynitrite (ONOO) downregulate endotoxin-stimulated prostaglandin release is the result of intracellular generation of ROS, including generation and availability of intracellular NO and ONOO and (ii) antioxidants and supranormal levels of intracellular NO and ONOO inhibit endotoxin-stimulated prostaglandin synthesis through effects on ecNOS and COX-2 mRNA. Both NO and ONOO have been generally thought to mediate oxidant tissue injury, but their excessive generation has recently been reported to confer both beneficial and deleterious effects. The proposed experiments represent a comprehensive approach to three specific aims and utilize techniques of cell biology, biochemistry and molecular biology. Experiments are proposed to determine: 1) the time course of induction of ecNOS and COX-2 following exposure to endotoxin and whether this is a consequence of intracellular oxidant stress, 2) the effects of exogenously generated NO and ONOO on the endotoxin response and whether the observed beneficial effects are the result of alterations in their intracellular level and, 3) whether endothelial cells genetically engineered to hyperexpress NO are protected from endotoxin-induced prostaglandin synthesis and release. Since the preliminary data also indicate differences in iNOS activity in bovine pulmonary artery endothelial cells (BPAEC) and bovine lung microvascular cells (BMVEC), the applicant will examine 1) and 2) in both BPAEC and BMVEC; human cells cultured from those same two sites will also be examined, as necessary. Improved understanding of the mechanism(s) of endotoxin-induced endothelial changes will result in improved and novel strategies for treatment of patients with the adult respiratory distress syndrome.

描述（改编自申请人的摘要）：申请人将 进一步观察细胞内反应性物质的产生 氧物质（ROS）有助于内毒素刺激的前列腺素 肺内皮的合成和释放。 两个密切相关的 根据初步数据提出假设，表明 内毒素导致细胞内产生ROS，并且抗氧化剂， DMSO，以及细胞外产生的一氧化氮 (NO) 和 过氧亚硝酸盐 (ONOO) 下调内毒素刺激的前列腺素释放 是细胞内ROS生成的结果，包括生成和 细胞内 NO 和 ONOO 的可用性以及 (ii) 抗氧化剂和 细胞内NO和ONOO的超正常水平抑制内毒素刺激 通过影响 ecNOS 和 COX-2 mRNA 来合成前列腺素。 两者都没有 和 ONOO 通常被认为介导氧化组织损伤，但是 最近有报道称，它们的过度生成赋予了两者 有益和有害的影响。 所提出的实验代表了 实现三个具体目标的综合方法并利用细胞技术 生物学、生物化学和分子生物学。 建议进行实验 确定： 1) ecNOS 和 COX-2 诱导的时间过程如下 接触内毒素以及这是否是细胞内毒素的结果 氧化应激，2）外源产生的NO和ONOO对 内毒素反应以及观察到的有益效果是否是 细胞内水平变化的结果，3) 是否 经过基因工程改造以超表达 NO 的内皮细胞受到保护 内毒素诱导的前列腺素合成和释放。 自从 初步数据还表明牛体内 iNOS 活性存在差异 肺动脉内皮细胞（BPAEC）和牛肺微血管 细胞（BMVEC），申请人将检查BPAEC和BMVEC中的1）和2）； 从这两个地点培养的人类细胞也将受到检查，因为 必要的。 加深对内毒素诱导机制的了解 内皮细胞的变化将导致改进和新颖的策略 成人呼吸窘迫综合征患者的治疗。