STEROID ACTION IN BACTERIAL ENDOTOXIC SHOCK

我们的假设是细菌内毒素能引起 来自网状内皮系统的介体 激素调节肝脏新陈代谢的变化。 在我们实验室的头21年半内获得的证据 该项目建议释放免疫调节性单细胞因子。 内毒素注射后与糖皮质激素相互作用 肌动蛋白基因表达。在这项研究的继续期间，我们 建议表征单一因子的作用，如IL-1，TNF， 和干扰素，在内毒素血症时糖皮质激素作用的改变。 这将通过滴定介体的抑制活性来实现- 丰富的血浆(从感染卡介苗的CF-1小鼠和 用2微克细菌内毒素攻击) 小鼠和鲁伯H35大鼠肝癌细胞(RHC)培养。更改后的 糖皮质激素对色氨酸加氧酶(TO)、酪氨酸的诱导 氨基转移酶(TAT)，磷酸烯醇式丙酮酸羧酸激酶 (PEPCK)和糖原合成酶(GS)将在 单核细胞治疗。这些肝酶的诱导受损 将与肝胞浆结合减少有关 地塞米松。此外，我们还将滴定其抑制活性 内毒素处理的库普弗细胞释放的因子。克罗斯- 富含介质的血浆和库普弗细胞液的吸附 针对已知的单因子的单特异性抗体将评估 单因子对血管内皮细胞可能的协同或拮抗作用 改变了糖皮质激素的作用。在拟议的研究中，我们还将 糖皮质激素对单核细胞介体的影响 被发现在内毒素过程中影响糖皮质激素的作用 令人震惊。我们将测量TO，TAT， 肾上腺切除(Adrx)小鼠和adrx小鼠的PEPCK和GS 用外源性类固醇重组。重组IL-1、肿瘤坏死因子和 干扰素将在小鼠和RHC中使用，以测试 类固醇对单核细胞的作用。这一提议的长期目标是 研究是为了了解潜在的分子事件 糖皮质激素-单核细胞因子的相互作用 内毒素休克时糖皮质激素的作用可能是 通过治疗干预逆转。这反过来将导致 提高了非常严重和危及生命的临床病例的存活率 综合症。

Our hypothesis is that bacterial endotoxin evokes the release of mediators from the reticuloendothelial system that mediate alterations in hormone regulated metabolism in the liver. Evidence obtained in our laboratory during the first 21/2 years of this project suggest immunomodulatory monokines, released following endotoxin administration interact with glucocorticoid actionon gene expression. In the continuation of this study we propose to characterize the role of monokines, such as IL 1, TNF, and IFN, in altered glucocorticoid action during endotoxemia. This will be done by titrating the inhibitory activity of mediator- rich plasma (obtained from CF-1 mice infected with BCG and challenged with 2 micrograms of bacterial endotoxin) in C3H/HeJ mice and Reuber H35 rat hepatoma cell (RHC) cultures. Altered glucocorticoid induction of tryptophan oxygenase (TO), tyrosine aminotransferase (TAT), phosphoenolpyruvate carboxykinase (PEPCK), and glyogen synthase (GS) will be monitored after monokine treatment. Impaired induction of these liver enzymes will be correlated with decreased hepatic cytosolic binding of dexamethasone. In addition we will titrate the inhibitory activity of factors released from endotoxin treated Kupffer cells. Cross- adsorption of mediator-rich plasma and Kupffer cell fluids with monospecific antibodies to known monokines will assess the possible synergistic or antagonistic effects of monokines on altered glucocorticoid action. In the proposed study we will also assess the influence of glucocorticoids on the monokine mediators that are found to affect glucocorticoid action during endotoxic shock. We will measure the inhibition of induction of TO, TAT, PEPCK and GS in adrenal-ectomized (adrx) mice and adrx mice reconstituted with exogenous steroid. Recombinant IL 1, TNF and IFN will be used in both mice and RHC to test the influence of steroids on monokine action. The long term goal of this proposed research is to understand the molecular events underlying glucocorticoid-monokine interactions so that the perturbation in glucocorticoid action during endotoxic shock may possibly be reversed with therapeutic intervention. This, in turn, will lead to improved survival from a very serious and life-threatening clinical syndrome.