ENDOTHELIAL CELL DERIVED NEUTROPHIL CHEMOATTRACTANTS

拟议研究的总体目标是调查 内皮细胞调节自身环境的潜力， 与肺血管损伤相关的疾病，肺血管损伤是导致 流动性和死亡率。 内皮细胞损伤与中性粒细胞 蓄积可能是这种疾病发病机制中的重要事件。 我们有初步证据表明接触组胺， 血管紧张素II、滑石粉或低氧环境刺激内皮细胞 以释放刺激中性粒细胞迁移的物质 在改良的Boyden室测定中。 环加氧酶的进一步研究 和脂氧合酶抑制剂，并使用反相高 高效液相色谱法（HPLC）表明，这些物质是 花生四烯酸的脂氧合酶代谢产物。 为了表征、纯化和 鉴定内皮细胞衍生的中性粒细胞化学引诱物活性， 我们将检查：1）热稳定性，2）在水溶液中的溶解度， 非水溶剂，3）对蛋白酶的稳定性，4）掺入的能力 3 H-花生四烯酸代谢物，6）反相进一步纯化 在初始HPLC上分离的化学引诱物活性峰的HPLC 7）对纯化后的样品进行扫描紫外光谱分析 与已知标准品的光谱相比， 来定义 与中性粒细胞化学引诱物产生相关的条件，我们 将确定：1）内皮细胞类型对生产的影响， 使用牛主动脉、牛肺动脉和人脐静脉 内皮细胞; 2）通过测试对内皮细胞的特异性 血管平滑肌细胞和IMR-90成纤维细胞，以及3） 内皮细胞单层的融合程度。 此外，本发明还提供了一种方法， 我们将比较内皮细胞衍生的中性粒细胞的性质， 由四种药剂-组胺，血管紧张素II， 滑石粉和缺氧-这可能通过不同的方式影响内皮细胞 手段，以提供洞察力的事件，可能引发的 与中性粒细胞化学引诱物的释放相关的代谢过程。 更好地理解内皮细胞调节其功能的能力， 通过这些研究获得的自身环境可能会导致更有效的 预防和治疗各种形式的血管损伤的方法。

The overall objective of the proposed research is to investigate the potential of the endothelial cell to regulate its own environment in conditions associated with lung vascular injury, an important cause of mobidity and mortality. Endothelial cell injury and neutrophil accumulation may be important events in the pathogenesis of this disorder. We have developed preliminary evidence that exposure to histamine, angiotensin II, talc, or a hypoxic environment stimulates endothelial cells in tissue culture to release substances that stimulate neutrophil migration in a modified Boyden chamber assay. Further studies with cyclo-oxygenase and lipoxygenase inhibitors and initial assays with reverse phase high performance liquid chromatography (HPLC) suggest that these substances are lipoxygenase metabolites of arachidonic acid. To characterize, purify and identify the endothelial cell derived neutrophil chemoattractant activity, we will examine: 1) heat stability, 2) solubility in aqueous or non-aqueous solvents, 3) stability to proteases, 4) ability to incorporate 3H-arachidonic acid metabolites, 6) further purification of reverse phase HPLC of peaks of chemoattractant activity resolved on the initial HPLC analysis, and 7) scanning ultraviolet spectrum analysis of the purified fractions as compared to the spectrum of known standards. To define the conditions associated with generation of neutrophil chemoattractant, we will determine: 1) the effect of endothelial cell type on production by using bovine aortic, bovine pulmonary arterial, and human umbilical vein endothelial cells; 2) the specificity for endothelial cells by testing vascular smooth muscle cells and IMR-90 fibroblasts, and 3) the effect of the degree of confluence of the endothelial cell monolayer. In addition, we will compare the nature of endothelial cell derived neutrophil chemoattractant induced by the four agents - histamine, angiotensin II, talc, and hypoxia - which likely influence endothelial cells by different means in order to provide insights into the events which might initiate the metabolic process associated with release of neutrophil chemoattractants. Better understanding of the ability of the endothelial cell to regulate its own environment obtained by these studies might lead to more effective methods of prevention and treatment of various forms of vascular injury.