REGULATION OF NFKB IN LUNG EPITHELIUM BY ROS/RNS

ROS/RNS 对肺上皮中 NFKB 的调控

• 批准号: 6125868
• 负责人: Yvonne M. W. Janssen-Heininger
• 金额: $ 23.73万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6125868
• 关键词: apoptosis; cell line; confocal scanning microscopy; free radicals; gene expression; hydrogen peroxide; inflammation; lung injury; mitogen activated protein kinase; nuclear factor kappa beta; oxidative stress; peroxynitrites; respiratory epithelium; transfection

The interaction of reactive nitrogen species (ROS/RNS) with lung epithelium, the first cell in contact with inhaled toxicants, may be critical in the initiation of pulmonary disease. Activation of signalling cascades and transcription factors may be important in determining the phenotypic outcome of exposure to these reactive metabolites and the activation of an inflammatory response. In this proposal we focus on the transcription factor, nuclear factor kappa B (Nf-kappaB) in a rat alveolar type II epithelial cell line (RLE) exposed to ROS or RNS. We hypothesize that activation of NF-kappaB by critical ROS/RNS occurs through unique signalling pathways that include mitogen- activated protein kinases (MAPK) and tyrosine kinases, and that the balance of these events is pivotal in the initiation of lung injury. NF-kappaB is critical in the regulation of genes with multiple functions that are involved in inflammation, immune immodulation or survival. Prevention of apoptosis by NF-kappaB may lead to activation of inflammatory genes, a hallmark of oxidant lung injury. Due to its' complex functions, the phenotypic implications of NF-kappaB activation in lung epithelium are obscure to date. In this proposal, we seek to investigate the mechanisms by which ROS or RNS activate NF-kappaB, the critical signalling pathways and the functional implications of NF- kappaB activation in RLE cells. We will investigate two critical RNS, hydrogen peroxide, or peroxynitrite to determine the implications of NF- kappaB activation and phenotypic endpoints (apoptosis and expression of inflammatory genes).Modulation of NF-kappaB by overexpression or prevention of its activation will allow us to determine the implications of NF-kappaB activation in lung epithelium by ROS and RNS. Furthermore, assessment of upstream signalling cascades and MAPKs as well as the modulation of various MAPKs will elucidate the critical signalling pathways by which ROS/RNS induce NF-kappaB. Combined approaches in specific aims 1-4 will lead to a better understanding of the role of NF- kappaB in pulmonary disease associated with exposure to ROS/RNS and may provide strategies for therapeutic intervention.

活性氮 (ROS/RNS) 与肺的相互作用 上皮细胞是第一个与吸入毒物接触的细胞 对于肺部疾病的发生至关重要。 激活 信号级联和转录因子可能在 确定暴露于这些反应性物质的表型结果 代谢物和炎症反应的激活。 在这个 建议我们关注转录因子，核因子kappa B (Nf-kappaB) 在大鼠肺泡 II 型上皮细胞系 (RLE) 中暴露 到 ROS 或 RNS。 我们假设 NF-κB 的激活通过关键 ROS/RNS 通过独特的信号通路发生，包括丝裂原- 激活蛋白激酶（MAPK）和酪氨酸激酶，并且 这些事件的平衡对于肺损伤的发生至关重要。 NF-kappaB 在多功能基因的调控中至关重要 与炎症、免疫调节或生存有关的物质。 NF-κB 预防细胞凋亡可能导致激活 炎症基因，氧化性肺损伤的标志。 由于其' 复杂功能，NF-kappaB 激活的表型影响 迄今为止，肺上皮细胞中的情况尚不清楚。 在本提案中，我们力求 研究 ROS 或 RNS 激活 NF-kappaB 的机制 关键信号通路和 NF-的功能意义 RLE 细胞中的 kappaB 激活。我们将研究两个关键的 RNS， 过氧化氢或过氧亚硝酸盐以确定 NF- 的影响 kappaB 激活和表型终点（细胞凋亡和表达 炎症基因）。通过过度表达或调节 NF-κB 防止其激活将使我们能够确定其影响 ROS 和 RNS 对肺上皮细胞中 NF-kappaB 的激活作用。此外， 上游信号级联和 MAPK 的评估以及 各种 MAPK 的调节将阐明关键信号传导 ROS/RNS 诱导 NF-κB 的途径。 组合方法 具体目标 1-4 将有助于更好地理解 NF-的作用 kappaB 与暴露于 ROS/RNS 相关的肺部疾病有关，并且可能 提供治疗干预策略。