EFFECTS ON AIRWAY PERMEABILITY AND CYTOSKELETON

对气道通透性和细胞骨架的影响

• 批准号: 3251260
• 负责人: Deepak K. Bhalla
• 金额: $ 17.97万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-03-01 至 1988-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3251260
• 关键词: aerosols; air sampling /monitoring; autoradiography; bronchus; cytochalasins; cytoskeleton; electrodes; electron microscopy; freeze etching; intercellular connection; membrane permeability; nitrogen oxides; organ culture; ozone; pollution related respiratory disorder; radiation detector; radiotracer; respiratory epithelium; respiratory gas transport; respiratory pharmacology; scintillation spectrometry; technetium; trachea

Cytoskeletal elements in the airway epithelial cells of the rat will be investigated as the possible intracellular target sites for 03 and N02. In vitro organ maintenance systems will be used to a) facilitate control of in vitro exposure to 03 (0.2 to 0.8 ppm) or N02 (0.5 to 5 ppm) or cytoskeleton-active agents; b) measure the effects of these agents on permeability and cytoskeletal structure; and c) compare effects of the agents with those produced by 03 and N02 in order to determine whether their mechanisms of action may involve the same or related cytoskeletal components. The observed effects will be related to the amount of each gas retained by tracheal tissue. In vitro experiments will be paralleled by selected in vivo studies in order to seek in vitro-in vivo correlations. Preliminary observations suggest that the mechanisms of action of cytoskeleton-active drugs such as cytochalasin B, which destabilizes the cytoskeleton and increases permeability, may be similar to that of 03 and N02, which also increase permeability. Cytoskeleton-stabilizing drugs, such as phalloidin and kinetin, may antagonize the effects of these gases. In vitro organ maintenance systems permit controlled exposure of rat tracheal tubes in measurement of quantity of gas retained. Transmucosal flux will be measured by applying tracer molecules (99mTc-DTPA, 3H-dextran, 125I-BSA) to the tracheal mucosa and sampling at the serosal surface for appearance of label. Immunocytochemistry by light and electron microscopy will be employed to characterize various cytoskeletal components and their relation to transport through tight junctions or endocytic vesicles. Freeze fracture will be employed in morphometry of tight junctions. The purpose of this study is to identify mechanisms of increased respiratory epithelial permeability upon exposure to 03 or N03. An added benefit may be the validation of an in vitro system for testing toxicity of gasses and aerosols. Such a system might be used for human respiratory tissue maintained and exposed in vitro to aid in extrapolating animal data to man in assessing risk of human exposure.

大鼠气道上皮细胞中的细胞骨架成分将被 作为O3和NO2的可能的细胞内靶位点进行研究。 在 体外器官维持系统将用于：a）促进对体内器官的控制， 体外暴露于O3（0.2至0.8ppm）或NO2（0.5至5 ppm）或 细胞增殖活性剂; B）测量这些试剂对 渗透性和细胞骨架结构;以及c）比较 与由O3和NO2产生的那些试剂进行比较，以确定是否 它们的作用机制可能涉及相同或相关的细胞骨架 件. 观察到的效果将与每种气体的量有关 由气管组织保留。 体外实验将由 选择体内研究，以寻求体外-体内相关性。 初步观察表明， 细胞增殖活性药物，如细胞松弛素B，它使细胞不稳定， 细胞骨架和增加渗透性，可能类似于03和 N 02，这也增加了渗透性。 细胞因子稳定药物， 例如鬼笔环肽和激动素，可以拮抗这些气体的作用。 体外器官维持系统允许大鼠受控暴露 用于测量保留气体量的气管导管。 经粘膜 通过应用示踪分子（99 mTc-DTPA，3 H-葡聚糖， 125 I-BSA）的气管粘膜，并在血清表面取样， 标签的外观 光镜和电镜免疫细胞化学 将用于表征各种细胞骨架成分及其 与通过紧密连接或内吞囊泡转运有关。 冷冻断裂将用于紧密连接的形态测定。 这项研究的目的是确定增加的机制， 在暴露于O3或NO3时的呼吸上皮通透性。 一个额外 有益之处可能是验证了用于测试药物毒性的体外系统。 气体和气溶胶。 这样的系统可以用于人类呼吸系统。 体外保存和暴露的组织，以帮助外推动物数据 评估人类接触的风险。

项目成果

Transport across rat trachea in vitro after exposure to cytoskeleton-active drugs in vitro or to ozone in vivo.

在体外接触细胞骨架活性药物或体内接触臭氧后，在体外通过大鼠气管转运。

• DOI: 10.3109/01902148909087857
• 发表时间: 1989
• 期刊: Experimental lung research
• 影响因子: 1.7
• 作者: Rasmussen,RE;Bhalla,DK
• 通讯作者: Bhalla,DK