Role of Eicosanoids In Modulating Endotoxin Induced Live

类二十烷酸在调节内毒素诱导的活体中的作用

• 批准号: 6577680
• 负责人: KENNETH L BRIGHAM
• 金额: $ 29.91万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6577680
• 关键词: adult respiratory distress syndrome; binding proteins; biological signal transduction; chimeric proteins; cytokine; eicosanoids; endotoxins; enzyme linked immunosorbent assay; gel mobility shift assay; gene expression; inflammation; lung injury; nuclear factor kappa beta; pathologic process; polymerase chain reaction; prostaglandin E; prostaglandin endoperoxide synthase; pulmonary edema; swine; transcription factor; transfection; vascular endothelium permeability; vascular resistance; vasoconstriction; western blottings

In the clinical setting of sepsis related ARDS, the presence of liver failure markedly increases mortality and studies of endotoxin induced lung injury in animals implicate effects on the liver as important for the full expression of the injury. The sequence of biochemical events that dictates the physiologic response to endotoxin may be activation of critical transcription factors, especially nuclear factor kappa B (NFkappaB) and CCAAT enhancer binding protein beta (C/EBPbeta, a.k.a. NF-IL6) in both the lungs and the liver, leading to generation of pro-inflammatory cytokines. Our preliminary data as well as data of others implicate eicosanoids as modulators of these endotoxin effects and induction of the cyclooxygenase (COX-the proximal enzyme in prostanoid synthesis) also involves NFkappaB and C/EBPbeta activation. To elucidate relationships among the physiologic, biochemical, molecular and pathophysiologic events related to liver-lung interactions in the endotoxin response we propose a series of studies in an in situ perfused swine model in which the lungs can be perfused with or without the liver in the perfusion circuit. We will: 1) determine effects of endotoxemia on pulmonary vascular resistance, lung vascular permeability, lung water content, tissue, perfusate and bronchoalveolar lavage fluid (BALF) concentrations of eicosanoids and cytokines, BALF total and differential cell counts, expression of TNFalpha, COX-1, COX- 2 and 5-lipoxygenase genes, activation of nuclear factor kappa B (NFkappaB) and C/EBPbeta in the lungs (and liver) with and without inclusion of the liver in the perfusion circuit; 2) manipulate concentrations of prostanoids in lung or liver by local controlled infusion of PGE2 into either organ, targeted delivery of a COX inhibitor to liver or lungs or transfecting the organ(s) with a construct expressing the COX gene and determine effects of these interactions on generation of eicosanoids and on endotoxin responses; 3) increase p20, the inhibitor isoform of C/EBPbeta, in the liver or the lungs by partial hepatic resection, delivery of a p20 membrane translocation signal fusion protein or transfection of the liver or lungs with a p20 gene and determine effects of these interventions on the endotoxin response; 4) inhibit activation of NFkappaB in liver or lungs by transfection of either organ with a gene encoding a transdominant inhibitor of NFkappaB activation or administration of a cell-permeable NFkappaB inhibitory fusion protein and determine effects on the endotoxin response. These studies will link pathophysiology to pathogenesis of the endotoxin response, providing a basis for identifying new potentially therapeutic interventions.

在脓毒症相关的ARDS的临床环境中，肝衰竭的存在显着增加了死亡率和内毒素诱导的动物肺损伤的研究，这对肝脏的影响对损伤的全部表达至关重要。决定对内毒素的生理反应的生物化事件的序列可能是关键转录因子的激活，尤其是核因子Kappa B（NFKappab）和CCAAT增强子结合蛋白β（C/EBPBETA，又称NF-IL6），肺和肝脏中的生产力。我们的初步数据以及其他数据都暗示类糖酸是这些内毒素作用的调节剂，并诱导环氧酶（Cox-the-the Prostanoid合成中的近端酶）也涉及NFKAPPAB和C/EBPBETA激活。为了阐明与内毒素反应中与肝脏肺相互作用有关的生理，生化，分子和病理生理事件之间的关系，我们提出了一系列在原位灌注猪模型中进行的一系列研究，其中可以在肺中灌注或不带有肝脏中的肝脏中灌注肺。我们将：1）确定内肌毒素对肺血管耐药性，血管渗透性，肺水含量，组织，灌注液和支气管肺泡灌洗液（BALF）eicosanoids和balf总细胞和差异细胞计数，Tnfalpha，cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1-2和cox-1-2和5 of Cox-1-of Cox，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1，Cox-1-2和5 Kappa B（NFKappab）和肺（和肝脏）中的C/EBPBETA，有和不包括肝脏灌注回路中； 2）通过将PGE2局部控制到器官中的局部控制的肺或肝脏中的前列腺素的浓度，靶向将Cox抑制剂送给肝脏或肺部或通过表达Cox基因的构建器的肝脏或转染器官，并确定这些相互作用的构造，并确定这些相互作用对生成毒素的影响以及对内毒素反应的影响； 3）增加p20，通过部分肝切除，肝脏或肺C/EBPBETA的抑制剂同工型，通过部分肝切除，P20膜易位信号融合蛋白或肝脏或肺部使用P20基因转染，并确定这些干预措施对内毒素反应的影响； 4）通过转染任何一种器官，用编码NFKAPPAB激活的跨剂量抑制剂或给予细胞可渗透的NFKAPAB抑制性融合蛋白并确定对内毒素反应的作用的基因，抑制NFKAPPAB在肝脏或肺中的激活。这些研究将将病理生理学与内毒素反应的发病机理联系起来，为鉴定新的潜在治疗干预措施提供了基础。