OXIDANTS AND ENDOTOXIN INDUCED ENDOTHELIAL INJURY

氧化剂和内毒素引起的内皮损伤

• 批准号: 2409244
• 负责人: BARBARA O MEYRICK
• 金额: $ 28.67万
• 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 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2409244
• 关键词: cell death; cyclic GMP; endotoxins; enzyme activity; enzyme induction /repression; glutamine; nitric oxide; nitric oxide synthase; nitrites; northern blottings; oxidative stress; peroxides; 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（根据申请人的摘要改编）：申请人将 进一步追求细胞内反应性的观察 氧（ROS）有助于内毒素刺激的前列腺素 合成和从肺部内皮释放。 两个密切相关 根据初步数据提出了假设，表明 内毒素会导致细胞内产生ROS，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的结果，包括产生和 细胞内NO和ONOO的可用性以及（ii）抗氧化剂和（ii） 细胞内NO和ONOO抑制内毒素刺激的超级水平 前列腺素通过对ECNOS和COX-2 mRNA的影响而合成。 两者都不 通常认为Onoo可以介导氧化组织损伤，但 最近据报道，他们过多的一代赋予了两者 有益和有害影响。 提出的实验代表 综合方法针对三个特定目标并利用细胞技术 生物学，生物化学和分子生物学。 提出了实验 确定：1）ECNOS和COX-2的诱导时间过程 暴露于内毒素，这是否是细胞内的结果 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作用是否是 其细胞内水平改变的结果，以及3）是否是否 内皮细胞基因设计为过度施加的NO受到保护 从内毒素诱导的前列腺素合成和释放。 自从 初步数据还表明牛的iNOS活性差异 肺动脉内皮细胞（BPAEC）和牛肺微血管 细胞（BMVEC），申请人将检查1）和2）在BPAEC和BMVEC中； 从相同两个地点培养的人类细胞也将被检查，因为 必要的。 对内毒素诱导的机制的理解有了改善 内皮变化将导致改进和新颖的策略 治疗成人呼吸窘迫综合征患者。