MODE OF ACTION OF STEROID HORMONES

类固醇激素的作用方式

• 批准号: 3224390
• 负责人: ALLAN U MUNCK
• 金额: $ 30.77万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-09-30 至 1990-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3224390
• 关键词: B lymphocyte; antibody receptor; antiinflammatory agents; chemical structure function; chromatography; colony stimulating factor; gene expression; glucocorticoids; glucose transport; hormone receptor; hormone regulation /control mechanism; immunosuppression; interferons; laboratory mouse; laboratory rat; macrophage; messenger RNA; monoclonal antibody; phosphorylation; radiotracer; thymus; tissue /cell culture

The long-term objective of this research is an understanding of actions of glucocorticoids on the immune system at molecular, cellular and physiological levels. Cells, tissues and functions of the immune system are major targets of glucocorticoids in their therapeutic functions as immunosuppressive and anti-inflammatory agents, and in the treatment of lymphocytic leukemias and lymphomas. Specific aims are directed towards three fundamental problems: (i) structural changes in glucocorticoid receptors that accompany "activation" (the transformation by which steroid-receptor complexes acquire affinity for nuclei and DNA); (ii) induction by glucocorticoids of mRNAs and proteins in rat thymus cells, in relation to rapid effects on glucose transport; (iii) interactions of glucocorticoids and gamma interferon on immune processes in intact mice. (i) The hypothesis that glucocorticoid receptors in the cell are cyclically phosphorylated and dephosphorylated, which emerged from earlier work on this project, will be tested by measuring 32P labeling of receptors in WEHI-7 mouse thymoma cells treated in various ways. Receptors purified with a monoclonal antibody will be analyzed for 32P content and location by peptide mapping, which will also be applied separately to identify peptides of the DNA-binding domain. The antibody will be used to identify nonbinding forms of receptors in energy-deprived cells. A kinetic model of receptor cycling will be tested with glucocorticoid antagonists. (ii) Glucocorticoids inhibit glucose transport in incubated rat thymus cells within 20 min, and exert other effects by 1 hour. Evidence suggests these effects are mediated by induced mRNAs coding for proteins that inhibit glucose transport, etc. Subtractive hybridization will be used to identify rare glucocorticoid-induced species of thymus mRNA and their respective proteins. Induced proteins will separately be sought by their putative ability to inhibit glucose transporters. (iii) Many potentially important effects of glycocorticoids on gamma interferon production and actions have been demonstrated in earlier work on this project with culture systems. Few have been demonstrated in animals. To bridge the gap between culture systems and whole animals, mice will be used to study the effects of adrenalectomy, exogenous gamma interferon and glycocorticoids on IgG-Fc receptor expression, B-cell (antibody) responses, and gamma interferon mRNA levels.

这项研究的长期目标是了解 糖皮质激素对免疫系统在分子，细胞和 生理水平。 免疫系统的细胞、组织和功能 是糖皮质激素治疗功能的主要靶点， 免疫抑制剂和抗炎剂，以及在治疗 淋巴细胞白血病和淋巴瘤。 具体目标针对三个基本问题： 糖皮质激素受体的结构变化伴随着“激活” (the类固醇-受体复合物获得亲和性的转化 对于细胞核和DNA）;（ii）通过糖皮质激素诱导mRNA， 大鼠胸腺细胞中的蛋白质，与对葡萄糖的快速作用有关 运输;（iii）糖皮质激素和γ干扰素对 完整小鼠的免疫过程。 (i)细胞中糖皮质激素受体呈周期性的假说 磷酸化和去磷酸化，这是从早期的工作中出现的， 这个项目，将通过测量受体的32 P标记进行测试， 以各种方式处理的WEHI-7小鼠胸腺瘤细胞。 纯化的受体 将通过以下方法分析32 P含量和位置： 肽图谱，也将单独应用于识别肽 DNA结合域。 抗体将用于识别 能量缺乏细胞中受体的非结合形式。 的动力学模型 将用糖皮质激素拮抗剂测试受体循环。 (ii)糖皮质激素抑制大鼠胸腺葡萄糖转运 细胞在20分钟内，并发挥其他作用的1小时。 证据表明 这些效应是由编码蛋白质的诱导mRNA介导的， 抑制葡萄糖转运等。消减杂交将用于 糖皮质激素诱导胸腺mRNA的罕见种类及其鉴定 各自的蛋白质。 诱导的蛋白质将分别寻找他们的 抑制葡萄糖转运蛋白的推定能力。 (iii)糖皮质激素对γ-淀粉样蛋白的许多潜在的重要作用 干扰素的产生和作用已经在早期关于 这个项目与文化系统。 很少有动物实验证明。 为了弥合培养系统和整个动物之间的差距， 用于研究肾上腺切除术、外源性γ干扰素和 糖皮质激素对IgG-Fc受体表达，B细胞（抗体）应答， 和γ干扰素mRNA水平。