LIPOXYGENASE PRODUCTS IN GLOMERULAR IMMUNE INJURY

肾小球内抗原-抗体复合物的存在是一种免疫反应。 在多种人类肾小球疾病中的常见启动机制。 虽然这些复合物的形成部位，它们形成的途径， 进入肾小球，它们的定位，以及肾小球的性质。 它们引起的组织病理学反应可能差异很大， 激活的白细胞在启动和维持致病性 事件是既定的。 PMN/巨噬细胞的主要后果 激活是释放有效的促炎氧化衍生物 花生四烯酸，特别是前列腺素类，血栓烷，和 白三烯 在过去的七年里，我们自己的工作和 其他研究者的研究已经建立了增强的肾小球合成， 有效的肾小球作用，以及5-和 15-花生四烯酸脂氧合酶（LO）产物在炎症过程中 大鼠肾小球损伤。 最近，我们获得了证据 肾小球内LO产物的合成可能不归因于 仅限于活化的白细胞，但涉及土著参与 肾小球细胞 这一点似乎特别重要， 这些细胞的跨细胞转化的关键 中性粒细胞/巨噬细胞衍生的代谢中间体，白三烯A4， 有效的促炎性类花生酸。 的表达分析 在正常和低浓度下， 病理条件可能提供重要的见解， 了解它们在肾小球病理生理学中的潜在作用。 利用培养的和新鲜分离的肾小球中mRNA的测量 细胞，原位杂交用于定量新鲜组织中的mRNA 切片，免疫细胞化学定位和Western印迹分析， 具体的酶，我们将研究细胞定位和时间 花生四烯酸脂氧合酶基因转录的调控， 翻译过程中肾小球免疫损伤。 我们将尝试 这些测量与最终产物合成速率的相关性， 重要的是，损伤的功能和组织形态学后遗症。 将研究肾小球疾病的两种代表性模型： 抗肾小球基底膜抗体诱导的肾小球炎，和 被动性Heymann肾炎 cDNA克隆的可用性和特异性 抗体的所有主要酶在这些途径，以及高度 灵敏和准确的检测分析技术， 他们的最终产品的定量，提供了一个独特的机会， 进行这种分析，有合理的成功机会。

The presence of antigen-antibody complexes within the glomerulus is a common initiating mechanism in a wide variety of human glomerulopathies. While the site of formation of these complexes, the route(s) by which they gain access to the glomerulus, their localization, and the nature of the histopathologic reaction they elicit can vary considerably, the central role of the activated leukocyte in initiating and perpetuating pathogenetic events is well-established. A major consequence of PMN/macrophage activation is the release of potent pro-inflammatory oxygenated derivatives of arachidonic acid, in particular prostanoids, thromboxanes, and leukotrienes. Over the past seven years, evidence from our own work and that of other investigators has established enhanced glomerular synthesis, potent glomerular actions, and pathophysiologic relevance for 5- and 15-lipoxygenase (LO) products of arachidonic acid during inflammatory glomerular injury in the rat. More recently, we have obtained evidence that the intraglomerular synthesis of LO products may not be attributed solely to activated leukocytes, but involve the participation of indigenous glomerular cells. This appears to be particularly relevant in the capacity of these cells to effect the transcellular transformation of a key neutrophil/macrophage-derived metabolic intermediate, leukotriene A4, into potent pro-inflammatory eicosanoids. Analysis of the expression of arachidonate LO pathway enzymes in glomerular cells under normal and pathologic conditions is likely to provide significant insight toward understanding their potential roles in glomerular pathophysiology. Utilizing measurements of mRNA in cultured and freshly isolated glomerular cells, in situ hybridization for quantitation of mRNA in fresh tissue sections, and immunocytochemical localization and Western blot analysis of specific enzymes, we will examine the cellular localization and temporal regulation of arachidonate lipoxygenase enzymes gene transcription and translation during the course of glomerular immune injury. We will attempt a correlation of these measurements with end-product synthetic rates and, importantly, with the functional and histomorphologic sequelae of injury. Two representative models of glomerular disease will be studied: anti-glomerular basement membrane antibody-induced glomerulitis, and passive Heymann nephritis. The availability of cDNA clones and specific antibodies for all the major enzymes in these pathways, as well as highly sensitive and accurate analytic techniques for the detection and quantitation of their end-products, provides a unique opportunity to undertake this analysis with a reasonable chance for success.