LIPOXYGENASE PRODUCTS IN GLOMERULAR IMMUNE INJURY

肾小球内抗原-抗体复合体的存在是一种 在多种人类肾小球疾病中共同的启动机制。 虽然这些复合体的形成地点，他们的路线(S)，他们 了解肾小球及其定位，以及肾小球的性质 它们引起的组织病理反应可能有很大不同，中央 激活的白细胞在致病和致病中的作用 活动由来已久。中性粒细胞/巨噬细胞的主要后果 活化就是释放强效的促炎含氧衍生物。 花生四烯酸，特别是前列腺素、血栓烷和 白三烯。在过去的七年里，来自我们自己的工作和 其他研究人员的研究已经证实了肾小球合成的增强， 强大的肾小球作用和5-和5-羟色胺的病理生理相关性 炎症过程中花生四烯酸的15-脂氧合酶(LO)产物 大鼠肾小球损伤模型。最近，我们获得了证据 LO产物在肾小球内的合成可能不是 仅限于激活的白细胞，但涉及土著参与 肾小球细胞。这似乎在能力上特别相关。 以影响关键基因的跨细胞转化 中性粒细胞/巨噬细胞衍生的代谢中间体白三烯A4进入 强效的促炎二十烷类化合物。试析《红楼梦》 正常和正常肾小球细胞花生四烯酸LO途径酶 病理情况很可能为 了解它们在肾小球病理生理学中的潜在作用。 培养肾小球和新鲜分离肾小球中mRNA的测定 细胞，原位杂交，定量新鲜组织中的信使核糖核酸 切片，免疫细胞化学定位和Western印迹分析 特定的酶，我们将检查细胞的定位和时间 花生四烯酸脂氧合酶基因转录与调控 肾小球免疫损伤过程中的翻译。我们将尝试 这些测量与最终产品合成速率的相关性，以及， 重要的是，伴随着损伤的功能和组织形态后遗症。 我们将研究两种具有代表性的肾小球疾病模型： 抗肾小球基底膜抗体诱导的肾小球炎，以及 被动型海曼肾炎。可获得的cDNA克隆和特异性 这些途径中所有主要酶的抗体，以及高度 灵敏、准确的检测和分析技术 他们最终产品的量化，提供了一个独特的机会 以合理的成功机会进行这项分析。

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.