MECHANISM OF ACTION OF STEROID HORMONES

我们正在调查分子和代谢事件涉及的 肾上腺糖皮质激素的几种不同作用的启动 模型系统提供了一些临床上更多的 重要的生物学效应：抑制炎症，抑制 免疫反应，细胞分化，以及抑制和 杀死胸腺淋巴细胞和淋巴癌细胞。平凡的 线程是我们的工作假设，即这些操作中的每一个都是由 通过激素诱导的改变(增加或减少)一个或多个 调节性蛋白质介体。在我们之前的工作中，我们开发了 通过生化研究发现这种媒介的证据，设计出新的 它们的检测手段(超高分辨率二维凝胶 )，并发现了一个或几个快速进化的 极低丰度快速翻转蛋白的诱导 (以及它们的mRNAs)在每种类型的靶细胞中；它们现在被识别 作为个人荷尔蒙作用的调解人的候选人。 在接下来的项目期内，我们很好地进入了第二阶段 研究，其中主要关注的是净化、开发 寡核苷酸和抗体探针，分子克隆， 这些蛋白的特性、亚细胞定位和调控 以及它们的mRNA。因为它们的丰度大多很低，有些 这项工作需要进一步制定创新战略，以 分离于二维凝胶上的次要蛋白的克隆。 随着全长DNA的获得，我们也进入了第三个阶段， 确定个别调解人的职能。这将是 通过构建和使用活病毒和其他 表达载体，以及通过其他方式。 我们已经发现、克隆并正在继续研究的一个介体是 可能具有特殊的临床相关性。这是一部小说 糖皮质激素调节的炎性环氧合酶(GRI-PGHS)，即 炎症显著增加，并显著抑制 糖皮质激素。它可能在发病机制中发挥核心作用。 许多折磨人类的炎症性疾病，如果不是所有的话， 糖皮质激素对其抑制作用提供了分子基础。 了解糖皮质激素最临床上有用的作用， 抑制炎症。

We are investigating the molecular and metabolic events involved in the initiation of several distinct adrenal glucocorticoid hormone actions in model systems that provide information about some of the more clinically important biological effects: suppression of inflammation, suppression of the immune response, cellular differentiation, and the suppression and killing of thymic lymphocytes and lymphoid cancer cells. The common thread is our working hypothesis that each of these actions is initiated through hormone-induced changes (increases or decreases) in one or more regulatory protein mediators. In our previous work we developed the evidence for such mediators through biochemical studies, devised novel means for their detection (ultra-high-resolution 2-dimensional gel electrophoresis), and discovered one or several rapidly-evolving inductions among the very-low-abundance rapidly-turning-over proteins (and their mRNAs) in each type of target cells; these are now recognized as candidates for mediators of the individual hormone actions. In the coming project period we are well into the second stage of this research, where the principal focus is on the purification, development of oligonucleotide and antibody probes, molecular cloning, characterization, sub-cellular location, and regulation of these proteins and their mRNAs. Since they are mostly of very low abundance some of this work requires the further development of innovative strategies for the cloning of the minor proteins resolved on 2-D gels. As the full-length cDNAs are obtained we are also entering a third stage, the determination of the functions for individual mediators. This will be accomplished by the construction and use of live virus and other expression vectors, and by other means. One mediator we have discovered, cloned, and are continuing to study is likely to have particular clinical relevance. It is a novel glucocorticoid-regulated inflammatory cyclooxygenase (gri-PGHS) that is dramatically increased in inflammation and remarkably suppressed by glucocorticoids. It is likely to play a central role in the pathogenesis in many if not all the inflammatory diseases that afflict mankind, and its suppression by glucocorticoids provides molecular basis for the understanding the most clinically useful of glucocorticoid actions, the suppression of inflammation.