ASBESTOS AND NO2 IN ENVIRONMENTAL LUNG DISEASE

环境性肺病中的石棉和二氧化氮

• 批准号: 2731250
• 负责人: NICHOLAS H HEINTZ
• 金额: $ 24.43万
• 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: 1998-08-15 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2731250
• 关键词: apoptosis; asbestos; autoradiography; biomarker; cell cycle; cell growth regulation; cell proliferation; cyclin dependent kinase; cytology; cytotoxicity; environmental toxicology; enzyme induction /repression; enzyme inhibitors; fibroblasts; flow cytometry; free radical oxygen; laboratory mouse; laboratory rat; lung injury; nitrogen oxides; nuclear factor kappa beta; occupational disease /disorder; respiratory epithelium; tissue /cell culture; transcription factor

DESCRIPTION Exposure of the human lung to environmental particulates and gases contributes to a variety of diseases, including pulmonary fibrosis and cancer. Previous work in our research group has shown that asbestos, reactive oxygen species (ROS), and reactive nitrogen species (RNS) trigger specific cell signaling pathways in target cells of the lung, leading to activation of transcription factors such as AP-1 and NF-KB. These and other transcription factors are involved in cellular decisions leading to phenotypic changes involved in the initiation of lung disease. To extend our understanding of cellular responses to mixtures of chemical and physical agents, we propose to study the effects of chrysotile asbestos and nitrogen dioxide (NO2), either alone or together, on three endpoints of exposure: cell survival, cell cycle progression, and apoptosis. First, we will characterize the responses of rat lung epithelial (RLE) cells and rat lung fibroblasts (RLF) to asbestos and/or N02, in regard to the activation of the transcription factors AP-1, NF-KB, and E2F and their downstream target genes. Activation of cyclin-dependent kinases (CDKs), metabolism of their protein inhibitors (i.e., CKIs p15, pl6, pl8, p2l, and p27),and phosphorylation of retinoblastoma family proteins (pRB and pl3O) will be studied as regulators of cell cycle progression. Cell imaging techniques, flow cytometry and activation of proteases will be used to measure apoptotic responses. Transient expression of dominantnegative regulatory molecules will be used to dissect mechanisms of exposure responses. For physiological relevance, results from experiments using models for cell cycle control and apoptosis in Vitro will be verified by inhalation studies using mice. Finally, to test the contributions of specific proteins such as p53, CKIS, and other candidate cell cycle regulators in cell cycle activation and/or apoptosis by asbestos and N02, inhalation studies will be performed in transgenic mice lacking specific genes of interest. Dissection of the role of cell cycle and survival regulators in proliferative and apoptotic responses to the combined effects of asbestos and N02 may lead to new biomarkers for exposure of the human lung to chemical mixtures.

描述 人肺暴露于环境颗粒物和气体 导致多种疾病，包括肺纤维化和 癌症。 我们研究小组之前的工作表明石棉， 活性氧 (ROS) 和活性氮 (RNS) 触发剂 肺部靶细胞中的特定细胞信号传导途径，导致 转录因子的激活，例如 AP-1 和 NF-KB。 这些和其他 转录因子参与细胞决策，导致 表型变化参与肺部疾病的发生。 延长 我们对细胞对化学和物理混合物反应的理解 剂，我们建议研究温石棉和氮的影响 二氧化氮（NO2），单独或一起，在三个暴露终点： 细胞存活、细胞周期进展和细胞凋亡。 首先，我们将 表征大鼠肺上皮 (RLE) 细胞和大鼠肺的反应 成纤维细胞 (RLF) 对石棉和/或 N02 的激活 转录因子 AP-1、NF-KB 和 E2F 及其下游靶标 基因。 细胞周期蛋白依赖性激酶 (CDK) 的激活及其代谢 蛋白质抑制剂（即 CKI p15、p16、p18、p2l 和 p27），以及 视网膜母细胞瘤家族蛋白（pRB 和 pl3O）的磷酸化将 作为细胞周期进程的调节剂进行研究。 细胞成像技术， 流式细胞术和蛋白酶激活将用于测量细胞凋亡 回应。 显性失活调节分子的瞬时表达 将用于剖析暴露反应的机制。 对于生理 相关性、使用细胞周期控制模型的实验结果以及 体外细胞凋亡将通过小鼠吸入研究来验证。 最后，测试特定蛋白质如p53、CKIS的贡献， 和细胞周期激活中的其他候选细胞周期调节剂和/或 石棉和 N02 引起的细胞凋亡，吸入研究将在 缺乏特定目的基因的转基因小鼠。 角色剖析 增殖和凋亡中细胞周期和存活调节因子的作用 对石棉和 N02 的综合影响的反应可能会导致新的 人肺暴露于化学混合物的生物标志物。