AUTOCRINE ROLE OF CYTOKINES IN VASCULAR SMOOTH MUSCLE

细胞因子在血管平滑肌中的自分泌作用

• 批准号: 2223770
• 负责人: Debbie Beasley
• 金额: $ 19.19万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2223770
• 关键词: autocrine; cell type; cytokine; endothelin; enzyme linked immunosorbent assay; human tissue; in situ hybridization; interleukin 6; laboratory rat; messenger RNA; nitric oxide; second messengers; tissue /cell culture; vascular smooth muscle; vasomotion

The long-range objective of this project is to determine the role of interleukin 1 (IL 1) and the related cytokines, interleukin 6 (IL 6), tumor necrosis factor (TNF), and interferon-gamma (IFN-gamma) in the physiological and pathophysiological modulation of vascular tone. The proposed research will test the hypotheses that autocrine or paracrine actions of IL1 and/or IL6 produced within blood vessels modulate systemic vascular resistance and local blood flow in vivo. These studies will: (1) Determine whether IL6, TNF, or IFN-gamma act independently or synergistically with IL1 to induce nitric oxide (NO) production in cultures human vascular smooth muscle cells (VSMC). (2) Determine whether cytokines, including IL6, TNF, and/or IL1, are produced by VSMC and mediate the known activation of NO synthase in VSMC by bacterial endotoxin. This will be achieved by analysis of released cytokines using enzyme-linked immunosorbent assays (ELISA), and by immunoneutralizations of cytokines using specific antibodies. (3) Determine which vascular cell types and vascular beds produce IL1 and IL6 in vivo, in both normal and LPS-treated rats, using in situ hybridization to detect IL1 and IL6 mRNA and immunocytochemistry to detect IL1 and IL6. (4) Determine whether second messengers or vasoactive agonists regulate IL1 or IL6 production and/or released by cultures human VSMC, using Norther analysis to measure IL1 and IL6 mRNA levels and ELISA techniques to measure IL1 and IL6. (5) Whether IL1 induces endothelin gene expression in human VSMC in vitro. The proposal is based on my recent demonstration that IL1, a monocyte- and macrophage-derived cytokine, is a vasodilator which acts directly on vascular smooth muscle. In contrast to the rapid and transient actions of endothelium-dependent vasodilators, the effect of IL1 is slow in onset and prolonged, lasting hours to days. Preliminary studies further indicate that IL1 and IL6 are produced by vascular tissue in vivo. Therefore, autocrine actions of IL2 and IL6 produced within VSMC may play a significant role in long-term modulation of vascular tone. Furthermore, cytokines produced by either immune cells or vascular cells probably contribute to the vasodilation associated with local inflammation, hemodialysis therapy, and septic shock, a major cause of human mortality. Understanding the roles of cytokines in modulating vascular tone may improve our ability to control these significant complications of human disease.

该项目的长期目标是确定 白细胞介素 1 (IL 1) 和相关细胞因子白细胞介素 6 (IL 6)、 肿瘤坏死因子（TNF）和干扰素-γ（IFN-γ） 血管张力的生理和病理生理调节。这 拟议的研究将检验自分泌或旁分泌的假设 调节血管内产生的 IL1 和/或 IL6 的作用 全身血管阻力和体内局部血流量。 这些研究将： (1) 确定IL6、TNF或IFN-gamma是否独立作用或 与 IL1 协同诱导一氧化氮 (NO) 的产生 培养人血管平滑肌细胞（VSMC）。 (2) 确定细胞因子，包括IL6、TNF和/或IL1是否是 由 VSMC 产生并介导 VSMC 中 NO 合酶的已知激活 由细菌内毒素引起。这将通过分析已发布的数据来实现 使用酶联免疫吸附测定 (ELISA) 检测细胞因子，并通过 使用特异性抗体对细胞因子进行免疫中和。 (3)确定哪些血管细胞类型和血管床产生IL1 和 IL6 体内，在正常和 LPS 处理的大鼠中，使用原位 杂交检测 IL1 和 IL6 mRNA，免疫细胞化学检测 检测IL1和IL6。 (4)确定第二信使或血管活性激动剂是否调节 IL1或IL6由培养物人VSMC产生和/或释放，使用 用于测量 IL1 和 IL6 mRNA 水平的 Norther 分析和 ELISA 技术 测量 IL1 和 IL6。 (5) IL1是否诱导人VSMC内皮素基因表达 体外。 该提案基于我最近的论证，即 IL1（一种单核细胞） 和巨噬细胞衍生的细胞因子，是一种血管舒张剂，直接作用于 血管平滑肌。与快速和短暂的动作相比 在内皮依赖性血管扩张剂中，IL1 的作用缓慢 起病时间长，持续数小时至数天。进一步的初步研究 表明IL1和IL6是由体内血管组织产生的。 因此，VSMC 内产生的 IL2 和 IL6 的自分泌作用可能 在血管张力的长期调节中发挥重要作用。 此外，免疫细胞或血管细胞产生的细胞因子 可能有助于局部血管舒张 炎症、血液透析治疗和感染性休克是导致感染的主要原因 人类死亡率。了解细胞因子在调节中的作用 血管张力可以提高我们控制这些重要的能力 人类疾病的并发症。