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Nuclear binding of glucocorticoid receptors. 糖皮质激素受体的核结合。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_8
S J Higgins, J D Baxter, G G Rousseau
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引用次数: 25
Glucocorticoids and hepatic glycogen metabolism. 糖皮质激素与肝糖原代谢。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_27
W Stalmans, M Laloux

The steady accumulation of glycogen in fetal rat liver during the last fifth of gestation is elicited by a transient rise in the level of circulating corticosterone. One effect of glucocorticoids is to induce glycogen synthase. The actual deposition of glycogen, however, depends on the appearance of a small amount of glycogen synthase in the active, dephosphorylated form. Induction of glycogen synthase phosphatase by glucocorticoids may explain the latter crucial process. Insulin enhances further the rate of glycogen deposition. The effect of insulin requires a previous exposure of the fetal liver to glucocorticoids. It is exerted on the enzyme interconversion system and appears not to involve new protein synthesis. Administration of glucocorticoids to adult fed or fasted animals causes within 3 h an intensive deposition of glycogen in the liver. This phenomenon is ultimately explained by both an activation of glycogen synthase and an inactivation of glycogen phosphorylase. The latter process may be due to an enhanced activity of phosphorylase phosphatase, or possibly of phosphorylase kinase phosphatase. The activation of glycogen synthase is explained by an enhanced activity of glycogen synthase phosphatase. The latter enzyme is normally profoundly inhibited by phosphorylase a; glucocorticoids cause the appearance in the liver of a protein factor that decreases and eventually cancels this inhibitory effect of phosphorylase a. It remains to be established whether or not some part of the glucocorticoid effect on adult liver is mediated by insulin.

在妊娠的最后五分之一,胎儿大鼠肝脏中糖原的稳定积累是由循环皮质酮水平的短暂上升引起的。糖皮质激素的作用之一是诱导糖原合成酶。然而,糖原的实际沉积依赖于少量活跃的、去磷酸化形式的糖原合成酶的出现。糖皮质激素诱导糖原合成酶磷酸酶可以解释后一个关键过程。胰岛素进一步提高糖原沉积速率。胰岛素的作用需要胎儿肝脏事先暴露于糖皮质激素。它作用于酶相互转化系统,似乎不涉及新的蛋白质合成。给喂食或禁食的成年动物注射糖皮质激素,会在3小时内引起肝糖原的大量沉积。这种现象的最终解释是糖原合成酶的激活和糖原磷酸化酶的失活。后一过程可能是由于磷酸化酶磷酸酶或磷酸化酶激酶磷酸酶活性增强所致。糖原合成酶的活化可以通过糖原合成酶磷酸酶活性的增强来解释。后一种酶通常被磷酸化酶a严重抑制;糖皮质激素导致肝脏中出现一种蛋白质因子,该蛋白因子降低并最终消除磷酸化酶a的抑制作用。糖皮质激素对成人肝脏的作用是否部分由胰岛素介导,尚不清楚。
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引用次数: 53
Mouse lymphoma cells: mechanisms of resistance to glucocorticoids. 小鼠淋巴瘤细胞:对糖皮质激素的抵抗机制。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_20
C H Sibley, K R Yamamoto

S49, an established line of mouse lymphoma cells, has several characteristics useful for the genetic analysis of glucocorticoid action: (1) a stable pseudodiploid karyotype; (2) an efficient cytolytic effect of glucocorticoids, which appears to follow the same biochemical pathway as steroid hormone action in other systems; (3) appearance of rare steroid-insensitive clones that exhibit stable, heritable resistance to further glucocorticoid treatment; (4) rapid growth in suspension culture and high cloning efficiency in soft agar, allowing facile isolation of variant clones. Two hundred individual steroid-resistant clones of S49 cells have been isolated and analyzed to determine the origin of their resistance. Most of these variant clones (55 %) fail to bind [3H]dexamethasone at levels above background; 70--75 percent bind less than 30 % of the wild-type level. The remaining clones fall into three general groups with respect to the efficiency with which receptors are translocated to the nucleus following dexamethasone treatment: one class transfers less than 10 percent of labeled receptors to the nucleus, another transfers normal amounts, and a third localizes virtually all of the receptors in the nucleus. The four variant phenotypes have been respectively designated r-, receptor activity deficient; nt-, nuclear transfer deficient; d-, deathless (appears normal in binding and nuclear transfer); and nti, increased nuclear transfer. Physical characterization by sucrose gradient sedimentation and gel permeation chromatography reveals that wild-type receptors are approximately 90,000 daltons and nti receptors about 50,000 daltons. The affinities of variant and wild-type receptors for purified DNA in vitro are consistent with their respective nuclear binding characteristics in vivo. Genetic studies with these and other cell lines, combined with recently developed methods for purification and structural analysis of minute quantities of proteins, can provide the level of biochemical resolution required for a fundamental understanding of the molecular mechanism of steroid hormone action.

S49是一种已建立的小鼠淋巴瘤细胞系,具有几个特征,可用于糖皮质激素作用的遗传分析:(1)稳定的假二倍体核型;(2)糖皮质激素的有效细胞溶解作用,其似乎遵循与其他系统中类固醇激素作用相同的生化途径;(3)出现罕见的类固醇不敏感克隆,对进一步的糖皮质激素治疗表现出稳定的、可遗传的抗性;(4)悬浮培养生长快,软琼脂克隆效率高,易分离变异克隆。已分离并分析了200个S49细胞的类固醇抗性克隆,以确定其抗性的来源。大多数这些变异克隆(55%)在高于背景水平时不能结合[3H]地塞米松;70% - 75%的人的结合水平不到野生型的30%。根据地塞米松治疗后受体转移到细胞核的效率,剩下的克隆分为三大类:一类将不到10%的标记受体转移到细胞核,另一类转移正常数量,第三类几乎将细胞核中的所有受体定位。这四种变异表型分别被命名为r-受体活性缺陷;Nt -,核转移缺陷;D -,不死(结合和核转移正常);核转移增加。通过蔗糖梯度沉降和凝胶渗透色谱的物理表征表明,野生型受体约为90,000道尔顿,反型受体约为50,000道尔顿。变异型和野生型受体在体外对纯化DNA的亲和力与其体内各自的核结合特性是一致的。对这些细胞系和其他细胞系进行遗传研究,结合最近开发的纯化和微量蛋白质结构分析方法,可以提供对类固醇激素作用的分子机制的基本理解所需的生化分辨率水平。
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引用次数: 18
Major Sex-Determining Genes 主要性别决定基因
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81261-3
Dr. Susumu Ohno
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引用次数: 207
Syneristic and antagonistic effects of glucocorticoids on insulin action. 糖皮质激素对胰岛素作用的协同和拮抗作用。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_32
T D Gelehrter

HTC cells, an established line of rat hepatoma cells in tissue culture, provide a useful experimental model system for studying the interaction of glucocorticoids and insulin in the regulation of protein metabolism. The actions of insulin and glucocorticoids on amino acid transport and protein degradation are antagonistic in this cell line. In contrast, the actions of these two hormones are additive with regard to the induction of tyrosine aminotransferase. The addition of insulin to HTC cells previously incubated with dexamethasone causes a rapid further doubling in the cellular concentration of this enzyme. The properties of the induction by insulin differ in several respects from the induction by glucocorticoids. The former occurs immediately, without the characteristic lag observed during induction by steroids. Insulin induction of transaminase does not require concomitant RNA synthesis, and does not cause the accumulation of specific mRNA for this enzyme as do glucocorticoids. Using specific immunoprecipitation techniques, we have demonstrated that insulin stimulates a nonselective increase in the rate of total protein synthesis in HTC cells, and a selective decrease in the rate of degradation of tyrosine aminotransferase relative to total protein. Thus the induction of transaminase by insulin involves two distinct actions of the hormone, affecting both synthesis and degradation of protein.

HTC细胞作为组织培养大鼠肝癌细胞系,为研究糖皮质激素和胰岛素相互作用调控蛋白质代谢提供了有益的实验模型系统。胰岛素和糖皮质激素对氨基酸转运和蛋白质降解的作用在该细胞系中是拮抗的。相反,这两种激素对酪氨酸转氨酶的诱导作用是加性的。将胰岛素添加到先前与地塞米松孵育的HTC细胞中,可使该酶的细胞浓度迅速进一步加倍。胰岛素诱导的性质在几个方面不同于糖皮质激素的诱导。前者立即发生,没有类固醇诱导时观察到的特征性滞后。胰岛素诱导转氨酶不需要同时合成RNA,也不像糖皮质激素那样引起转氨酶特异性mRNA的积累。使用特异性免疫沉淀技术,我们已经证明胰岛素刺激HTC细胞中总蛋白合成速率的非选择性增加,以及酪氨酸氨基转移酶相对于总蛋白的降解速率的选择性降低。因此胰岛素对转氨酶的诱导涉及激素的两种不同的作用,影响蛋白质的合成和降解。
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引用次数: 8
Allosteric and competitive steroid-receptor interactions. 变构和竞争性类固醇受体相互作用。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_7
M R Sherman
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引用次数: 12
Glucocorticoid hormone action: an overview. 糖皮质激素的作用综述。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_1
J D Baxter, G G Rousseau

An overview of glucocorticoid action is presented including aspects of historic, clinical, and physiologic interest. Despite the diversity of effects, glucocorticoid hormone action at the cellular level simplifies to a rather universal model that involves binding to a soluble receptor, followed by interaction of the complex with the chromatin and modification of gene expression. This unitary concept has important implications in pathology, pharmacology, and therapeutics. Finally, a definition of a glucocorticoid in terms of its receptor is presented, which we feel is better than the traditional terminology based on specific biological effects. It is hoped that this review will also provide the reader with a framework in which the various contributions in the Monograph can be integrated.

概述糖皮质激素的作用,包括历史,临床和生理方面的兴趣。尽管作用多种多样,但糖皮质激素在细胞水平上的作用简化为一种相当普遍的模式,即与可溶性受体结合,随后与染色质相互作用,并修饰基因表达。这个统一的概念在病理学、药理学和治疗学中具有重要意义。最后,提出了糖皮质激素受体的定义,我们认为这比基于特定生物效应的传统术语更好。希望这篇评论也将为读者提供一个框架,在这个框架中,专著中的各种贡献可以整合在一起。
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引用次数: 157
Regulation of growth hormone messenger RNA. 生长激素信使RNA的调控。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_15
J A Martial, P H Seeburg, D T Matulich, H M Goodman, J D Baxter

In cultured rat pituitary cells, glucocorticoids regulate growth hormone production by modulating the number of growth hormone messenger RNA molecules. The effect is quite specific, since only a few other mRNAs are affected by the hormones. This response is demonstrated by assays involving cell-free mRNA translation and cDNA-RNA hybridization. Furthermore, the inducibility by the glucocorticoids is regulated by at least one other class of hormones, thyroid hormone. Thus, this system serves as a model for studying not only the glucocorticoid regulation of specific mRNA, but also the control of this regulation by other factors in the target tissue.

在培养的大鼠垂体细胞中,糖皮质激素通过调节生长激素信使RNA分子的数量来调节生长激素的产生。这种影响是非常特殊的,因为只有少数其他mrna受到激素的影响。这种反应是通过无细胞mRNA翻译和cDNA-RNA杂交实验证明的。此外,糖皮质激素的诱导作用受至少另一类激素(甲状腺激素)的调节。因此,该系统不仅可以作为研究糖皮质激素对特定mRNA调控的模型,还可以作为研究靶组织中其他因素对这种调控的控制的模型。
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引用次数: 11
Effects of glucocorticoids on fibroblasts. 糖皮质激素对成纤维细胞的影响。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_18
L Aronow

Mouse fibroblasts growing in vitro respond to glucocorticoids in a dose-dependent fashion by reduced rates of growth. The inhibition of growth observed in vitro is related to the topical anti-inflammatory action of glucocorticoids and to their capacity to inhibit wound repair. The cells growing in vitro possess a glucocorticoid receptor system that has been studied in some detail using [3H]triamcinolone acetonide as a radiolabeled ligand. The initial binding reaction occurs in the cytosol. The complex is then rapidly taken up in the nucleus of the cell by a temperature-sensitive process. In the nucleus, the complex exists in two forms, one of which is readily extracted by 0.3 M KCl solutions. A small amount of steroid-receptor complex is tightly bound to chromatin. Under normal incubation conditions, there is a constant cycling of steroid-receptor complex, and unbound receptor is generated back into the cytosol from the nucleus with a half-life of about 30 min. Regeneration of unbound receptor does not depend on protein synthesis and is a temperature-sensitive and energy-requiring process. Incubating the steroid-treated cells in the absence of glucose and in the presence of inhibitors such as cyanide or dinitrophenol leads to a loss of cytoplasmic steroid-receptor complexes, and an accumulation of the complex in the nuclear residual form, tightly bound to chromatin. With respect to nuclear effects of steroid treatment, we have found that incubating fibroblasts in vitro with glucocorticoids produces a prompt decrease in the amount of a satellite H1 histone found in these cells.

体外生长的小鼠成纤维细胞对糖皮质激素的反应以剂量依赖性的方式降低生长速率。体外观察到的生长抑制与糖皮质激素的局部抗炎作用及其抑制伤口修复的能力有关。体外生长的细胞具有糖皮质激素受体系统,使用[3H]曲安奈德作为放射性标记配体对其进行了一些详细的研究。最初的结合反应发生在细胞质溶胶中。然后,通过对温度敏感的过程,这种复合物被迅速地吸收到细胞核中。在细胞核中,络合物以两种形式存在,其中一种很容易被0.3 M的氯化钾溶液提取。少量的类固醇受体复合体与染色质紧密结合。在正常孵育条件下,类固醇受体复合物不断循环,未结合受体从细胞核生成回细胞质,半衰期约为30分钟。未结合受体的再生不依赖于蛋白质合成,是一个对温度敏感和需要能量的过程。在没有葡萄糖和抑制剂(如氰化物或二硝基酚)存在的情况下培养类固醇处理过的细胞,导致细胞质类固醇受体复合物的损失,并以核残留形式积累复合物,紧密结合于染色质。关于类固醇治疗的核效应,我们发现在体外用糖皮质激素培养成纤维细胞会迅速减少这些细胞中发现的卫星H1组蛋白的数量。
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引用次数: 19
Inhibition of glucose transport in fat cells and activation of lipolysis by glucocorticoids. 糖皮质激素对脂肪细胞中葡萄糖转运的抑制和对脂肪分解的激活。
Pub Date : 1979-01-01 DOI: 10.1007/978-3-642-81265-1_29
J N Fain

The major effects of glucocorticoids on white fat are shown in Fig. 1. The glucocorticoid diffuses into the cytosol of fat cells where it binds to a soluble receptor. The steroid-receptor complex then enters the nucleus where RNA synthesis is increased. The next step may be a selective increase in synthesis of protein(s). In any event, there is an inhibition of the membrane-bound glucose transport system and an increase in the ability of lipolytic agents to activate triglyceride lipolysis. There is also a decrease in phosphoenolpyruvate carboxykinase, lipoprotein lipase, and fatty acid synthetase that occurs after a somewhat longer lag period than is required for inhibition of glucose transport or lipolysis activation. Whether these effects are independent or secondary to the glucose transport inhibition and lipolysis activation remains to be established.

糖皮质激素对白色脂肪的主要作用如图1所示。糖皮质激素扩散到脂肪细胞的细胞质中,在那里它与可溶性受体结合。然后类固醇受体复合物进入细胞核,在那里RNA合成增加。下一步可能是选择性地增加蛋白质的合成。在任何情况下,膜结合葡萄糖转运系统受到抑制,脂溶剂激活甘油三酯脂溶的能力增强。磷酸烯醇丙酮酸羧激酶、脂蛋白脂肪酶和脂肪酸合成酶的减少也发生在比抑制葡萄糖转运或脂解激活所需的较长滞后期之后。这些作用是独立的还是继发于葡萄糖转运抑制和脂解激活还有待确定。
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引用次数: 33
期刊
Monographs on endocrinology
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