STEROID ACTION IN BACTERIAL ENDOTOXIC SHOCK

细菌内毒素休克中的类固醇作用

• 批准号: 3129901
• 负责人: RODERICK E MCCALLUM
• 金额: $ 4.51万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-03-01 至 1987-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3129901
• 关键词: Escherichia coli; Kupffer's cell; adrenalectomy; carbohydrate metabolism; cell free system; dexamethasone; endotoxins; glucocorticoids; gluconeogenesis; gram negative bacteria; hormone regulation /control mechanism; host organism interaction; hypoglycemia; kidney; liver cells; liver metabolism; molecular pathology; spleen; steroid hormone metabolism; striated muscles

Bacterial endotoxins (ETX), components of the cell wall of gram-negative bacteria, elicit a number of cardiovascular, inflammatory, and hormonal responses. ETX-mediated alterations in metabolism, including glycogen depletion and diminished gluconeogenesis, occur at a time when plasma glucocorticoid, glucogan and epinephrine levels are normal to elevated. Our overall objective is to determine the mechanism(s) by which ETX perturbs host carbohydrate metabolism during gram-negative speticemia. This study is based on the hypothesis that ETX interferes with the normal action of glucocorticoids at the molecular level. To examine this hypothesis, we propose to investigate the consequences of down regulation of glucocorticoid receptors during endotoxemia. We will extend preliminary studies by comparing glucocorticoid binding to liver, spleen, skeletal muscle, and kidney cytosolic receptors from ETX-treated mice. We will determine whether the effects of ETX on cytosolic binding are direct or mediated by adding ETX directly to isolated normal hepatocytes in vitro, and with the passive transfer of serum and peritoneal cells between ETX-normal responder mice and ETX-low responder mice. Soluble mediators that may regulate hepatic glucocorticoid receptors will be screened for, using cultured peritoneal exudate cells and isolated Kupffer cells. Purification and characterization of soluble factors found to regulate steroid receptors will be pursued. A secondary aim of this study is to examine other aspects of intracellular steroid action in liver during endotoxic shock. We will determine the magnitude of nuclear binding of steroid-protein complexes in liver (in vivo) and in hepatocyte (in vitro) preparations. RNA polymerase activity will be measured, as will the ability of mRNA isolated from endotoxin-treated and control mouse livers to direct protein synthesis in a cell-free translation system. Successful completion of this study will yield data describing whether interference with glucocorticoid action in liver ETX is direct or mediated. In addition, the project will ultimately lead to an understanding of the role of altered hepatic metabolism in bacterial sepsis and shock.

细菌内毒素（ETX），革兰氏阴性菌细胞壁组分 细菌，引发许多心血管，炎症和激素 应答 ETX介导的代谢改变，包括糖原 消耗和减少的再生，发生在当血浆 糖皮质激素、胰高血糖素和肾上腺素水平正常到升高。 我们的总体目标是确定ETX 在革兰氏阴性菌血症期间干扰宿主的碳水化合物代谢。 本研究基于ETX干扰正常 糖皮质激素在分子水平上的作用。 审查这个 假设，我们建议调查下调的后果 糖皮质激素受体的变化 我们将延长初步 通过比较糖皮质激素与肝脏、脾脏、骨骼 肌肉和肾胞质受体。 我们将 确定ETX对胞质结合的影响是直接的还是 通过将ETX直接加入体外分离的正常肝细胞介导， 并且随着血清和腹膜细胞在 ETX正常应答小鼠和ETX低应答小鼠。 可溶性介质 可能调节肝脏糖皮质激素受体的基因将被筛选， 使用培养的腹膜渗出细胞和分离的枯否细胞。 可溶性调节因子的纯化和表征 将追踪类固醇受体。 本研究的第二个目的是 检查其他方面的细胞内类固醇作用在肝脏中， 内毒素休克 我们将确定核约束力的大小， 肝脏（体内）和肝细胞（体外）中类固醇-蛋白质复合物 准备工作。 将测量RNA聚合酶活性， 从内毒素处理的和对照小鼠肝脏中分离的mRNA的能力， 在无细胞翻译系统中直接合成蛋白质。 成功 完成这项研究将产生数据，描述是否干扰 与糖皮质激素作用在肝脏ETX是直接或介导的。 在 此外，该项目将最终导致对角色的理解 在细菌性败血症和休克中改变肝脏代谢。