CYCLOOXYGENASE IN GLOMERULAR ENDOTHELIAL AND EPITHELIAL CELLS

肾小球内皮细胞和上皮细胞中的环加氧酶

• 批准号: 3733110
• 负责人: Richard Hoover
• 金额: --
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3733110
• 关键词: aspirin; biological signal transduction; cell cell interaction; cell growth regulation; chronic renal failure; complement pathway; eicosanoid metabolism; enzyme activity; enzyme inhibitors; epithelium; extracellular matrix proteins; fluorescence microscopy; glomerulonephritis; interleukin 1; laboratory rat; pathologic process; platelet derived growth factor; prostaglandin endoperoxide synthase; renal glomerulus; thrombin; tissue /cell culture; vascular endothelium

End stage renal disease producing proteinuria and glomerular sclerosis can be initiated by both immune, e.g., inflammation, and non-immune, e.g., hypertension, injuries. The early events in these injuries are associated with changes in the individual cells of the glomeruli. Whether there is a common mechanism through which these injuries lead to progressive nephron destruction is the basis of this project. We hypothesize that both types of injuries affect glomerular synthesis of arachidonic acid metabolites by the cyclooxygenase pathway and these metabolites contribute to progression. We propose to examine in vitro the effects of immune and non-immune perturbations on the cyclooxygenase pathway in glomerular endothelial and epithelial cells. Specifically, we want (1) to characterize and quantitate the cyclooxygenase products of these cells as a result of conditions which mimic immune (thrombin, IL-1, PDGF and complement activation) and non- immune (stretching/relaxation) injuries; (2) to determine the effects of these conditions on the synthesis and activation of the cyclooxygenase enzyme; (3) to investigate the parameters of enzyme recovery in both cells after irreversible inhibition with aspirin; (4) to evaluate the effect f arachidonate metabolites produced by one cell on another cell; (5) to determine if the glomerular cells communicate with one another through these cyclooxygenase products and if this could be a form of self regulation of cyclooxygenase activation; (6) to examine the signal transduction mechanisms involved with activation of cyclooxygenase and those activated by cyclooxygenase products; and (7) to determine if another endothelial cell located outside the glomerulus, i.e., the endothelial cell of the afferent arteriole displays heterogeneity with respect to cyclooxygenase metabolites. These experiments will require the use of several different experimental methods, including cell culture techniques, biochemical analyses of the cyclooxygenase enzyme and its products, analyses of extracellular matrix components, growth studies, morphological analyses, RNA analyses, permeability studies, fluorescent techniques, and biochemical analyses of intracellular events. This project will rely upon the expertise of the three Cores in the Center Grant: B - the analytical core (quantitative and structural analysis of cyclooxygenase metabolites); C - the morphology and imaging core; and D - the extracellular matrix analysis core. These experiments are all in vitro and are designed to complement the in vivo studies in Projects 1 and 2. It is hoped that the results of these experiments will help in devising interventions to reduce the impact of these injuries and prevent progressive nephron destruction.

产生蛋白尿和肾小球硬化症的末期肾脏疾病可以 可以通过免疫，例如炎症和非免疫来引发 高血压，受伤。 这些伤害的早期事件是相关的 随着肾小球单个细胞的变化。 是否有一个 这些伤害导致渐进性肾单位的常见机制 破坏是该项目的基础。 我们假设两种类型 损伤会影响奏肾合成蛛网膜酸代谢产物的肾小球合成 环氧合酶途径和这些代谢产物有助于进展。 我们建议在体外检查免疫和非免疫的影响 肾小球内皮和环氧化酶途径的扰动 上皮细胞。 具体来说，我们希望（1）表征和定量 由于条件，这些细胞的环氧合酶产物 模拟免疫（凝血酶，IL-1，PDGF和补体激活）和非 - 免疫（拉伸/放松）伤害； （2）确定 这些条件关于环氧合酶的合成和激活 酶； （3）研究两个细胞中酶恢复的参数 对阿司匹林不可逆抑制后； （4）评估效果f 一个细胞在另一个细胞上产生的蛛形代谢产物； （5）到 确定肾小球细胞是否通过 这些环氧合酶产品，如果这可能是一种自我形式 环氧合酶激活的调节； （6）检查信号 与环氧合酶激活有关的转导机制和 那些被环氧酶产物激活的人； （7）确定是否另一个 内皮细胞位于肾小球外，即内皮细胞 传入小动物的表现相对于 环氧合酶代谢产物。 这些实验将需要使用 几种不同的实验方法，包括细胞培养技术， 环氧合酶及其产物的生化分析， 细胞外基质成分，生长研究，形态学的分析 分析，RNA分析，渗透率研究，荧光技术和 细胞内事件的生化分析。 这个项目将依靠 中心赠款中三个核心的专业知识：B-分析 核心（环氧酶代谢产物的定量和结构分析）； C-形态和成像核心；和D-细胞外基质 分析核心。 这些实验全都在体外，设计为 补充项目1和2中的体内研究。希望 这些实验的结果将有助于制定干预措施以减少 这些伤害的影响并防止进行性肾脏毁灭。