Interleukin-4 effects on calcium in airway smooth muscle

Interleukin-4 对气道平滑肌钙的影响

• 批准号: 6433927
• 负责人: JOHN Mark MADISON
• 金额: $ 35.78万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-04-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6433927
• 关键词: androstane compound; animal tissue; beta adrenergic receptor; bronchomotion; calcium channel; calcium flux; calcium indicator; calcium transporting ATPase; cell line; cell morphology; electrophysiology; inositol phosphates; interleukin 4; muscle cells; protein tyrosine kinase; receptor coupling; respiratory muscles; sarcoplasmic reticulum; smooth muscle; voltage gated channel

DESCRIPTION (provided by applicant): The long-term objectives have been to understand calcium regulation in airway smooth muscle. Intracellular calcium stores of the sarcoplasmic reticulum (SR) play a role in determining the cytosolic calcium concentrations ([Ca2+]i) that regulate smooth muscle functions, including contraction. Therefore, understanding the mechanisms by which stimuli cause release of calcium from the SR to generate a rapid increase in [Ca2+]i in the cell cytosol (calcium transient) and how the SR then refills with calcium is important. Work on SR refilling has led to the novel observation that interleukin (IL)-4 rapidly (<30 min) inhibits carbachol-stimulated calcium transients in bovine airway smooth muscle cells. This new finding is potentially important because it conceptually links calcium regulation in airway smooth muscle cells to the T helper 2-type cytokine profile characteristic of asthmatic airway inflammation. Notably, IL-4 is closely related to IL-l3, an important mediator of airway inflammation, and binds to a shared family of receptor complexes. In general, the aims are to determine how IL-4 rapidly inhibits calcium transients and to assess for effects on contraction. Specifically, the goals are to determine the effects of IL-4 on 1) SR calcium content; 2) calcium influx; and 3) coupling of receptors to inositol trisphosphate generation. Then, the goals are to 4) characterize signal transduction pathways underlying IL-4 effects on calcium and 5) characterize the effects of IL-4 on cell shortening. The principal methods are single cell, ratiometric imaging of [Ca2+]i, including three dimensional reconstruction of images; electrophysiology to assess calcium influx; and biochemistry to define signal transduction pathways. Studies will be done with freshly-dispersed bovine and cultured human airway smooth muscle cells.

描述（申请人提供）：长期目标是 了解气道平滑肌的钙调节。细胞内钙 肌质网（SR）的存储在确定 调节平滑肌的胞质钙浓度（[Ca2+] i） 功能，包括收缩。因此，了解机制 哪些刺激导致钙从SR释放，从而迅速增加 在细胞细胞质（钙瞬态）中的[Ca2+] i中，然后如何补充SR 使用钙很重要。 SR补充的工作导致了小说 观察白介素（IL）-4快速抑制（<30分钟） 卡尔巴乔刺激的牛气道平滑肌细胞中的钙瞬变。 这个新发现可能很重要，因为它从概念上联系了钙 气道平滑肌细胞中的调节对T辅助2型细胞因子的调节 哮喘气道炎症的特征。值得注意的是，IL-4是 与IL-L3密切相关，IL-L3是气道炎症的重要介体， 结合共享的受体复合物家族。通常，目的是 确定IL-4如何迅速抑制钙瞬变并评估 对收缩的影响。具体而言，目标是确定 IL-4 ON 1）SR钙含量； 2）钙涌入； 3）受体耦合 至肌醇三磷酸盐的产生。然后，目标是4）特征 IL-4对钙的影响的信号转导途径和5） 表征IL-4对细胞缩短的影响。主要方法是 [Ca2+] i的单细胞，比率成像，包括三维 图像的重建；电生理学评估钙涌入；和 生物化学以定义信号转导途径。研究将进行 新鲜分散的牛和培养的人类气道平滑肌细胞。