REGULATION OF E-SELECTIN DURING ACUTE INFLAMMATION

急性炎症期间 E-选择素的调节

• 批准号: 2634278
• 负责人: KEVIN Lenuell BILLUPS
• 金额: $ 10.34万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2634278
• 关键词: RNase protection assay; SDS polyacrylamide gel electrophoresis; acute disease /disorder; enzyme linked immunosorbent assay; gene expression; genetic regulation; heart cell; immunocytochemistry; inflammation; ischemia; kidney cell; laboratory rat; lipopolysaccharides; messenger RNA; monoclonal antibody; northern blottings; nucleic acid sequence; phenotype; polymerase chain reaction; reperfusion; selectins; synthetic peptide; vascular endothelium; western blottings

Acute inflammation is a major cause of morbidity and mortality in medicine today. Many common disorders of the urogenital tract involve some aspect of acute or chronic inflammation. Examples of specific disorders include: pyelonephritis, cystitis, epididymitis, orchitis, and prostatitis. In addition to acute infectious etiologies, it is becoming increasingly clear that ischemia/reperfusion injury triggers a striking acute inflammatory response that contributes to tissue damage and organ failure. Examples is ischemic injury in the urogenital tract include renal transplantation, trauma, parenchymal sparing surgery for cancer, and torsion of the spermatic cord. I hypothesize that one of the earliest events to occur during an acute inflammatory response is the expression of endothelial cell adhesion molecules. This proposition is supported by a large body of in vitro and in vivo data examining endothelial cell activation. E- selectin is unique among adhesion molecules because it is found only on activated endothelial cells. B-selectin has been shown to be an important regulator of early leukocyte adhesion. Understanding the role of E- selectin in vivo has been difficult primarily because a lack of bioactive molecular probes and antibodies, especially for use animal models. I propose to use the rat kidney and heart as model tissues to study in vivo expression of E-selectin during acute inflammation induced by injection of lipopolysaccharide or ischemia/reperfusion injury. Initial studies will focus on a detailed characterization of E-selectin mRNA expression, using recently cloned rat E-selectin cDNA fragments. Novel studies, using RNase protection assays, will be performed to investigate preliminary evidence for the existence of E-selectin mRNA splice variants. Another objective of these experiments is to generate antibodies to characterize protein expression of E-selectin during acute inflammation. The time course and phenotype of infiltrating leukocytes will be studied, using immunocytochemistry, and these data compared to the time course of E- selectin gene expression. The goal of this project is to gain insight into cellular and molecular mechanisms regulating expression of E- selectin. Long term objectives include applying information on these regulatory mechanisms to the study of acute and chronic inflammatory disorders of the urogenital tract. To the best of my knowledge no other laboratory possesses the molecular tools to investigate such novel aspects of E-selectin, in vivo. Data obtained from these proposed studies would impact on a number of conditions affecting urogenital organs and set the stage for later investigations focus on chronic inflammatory disorders (i.e. interstitial cystitis, prostatitis, inflammatory aspects of prostatic hypertrophy).

急性炎症是医学发病率和死亡率的主要原因 今天。泌尿生殖道的许多常见疾病都涉及某些方面 急性或慢性炎症。 特定疾病的示例包括： 肾盂肾炎，膀胱炎，附睾炎，提供卵形和前列腺炎。在 除了急性感染病因，它变得越来越清楚 缺血/再灌注损伤会触发引人注目的急性炎症 导致组织损伤和器官衰竭的反应。示例是 泌尿生殖道缺血性损伤包括肾移植， 创伤，实质性释放手术的癌症和扭转 精子线。我假设最早发生的事件之一 在急性炎症反应中是内皮的表达 细胞粘附分子。这个主张得到了一大批主体的支持 体外和体内数据检查内皮细胞激活。 e- 选择素在粘附分子中是独一无二的 活化的内皮细胞。 B-选择素已被证明是重要的 早期白细胞粘附的调节剂。了解电子的作用 体内selectin很难主要是因为缺乏生物活性 分子探针和抗体，特别是用于使用动物模型。我 建议将大鼠肾脏和心脏作为模型组织在体内研究 通过注射诱导的急性炎症期间的E-选择蛋白表达 脂多糖或缺血/再灌注损伤。最初的研究将 专注于使用E-选择蛋白mRNA表达的详细表征 最近克隆的大鼠e-选择蛋白cDNA片段。新研究，使用RNase 保护测定法将进行调查初步证据 为了存在E-选择蛋白mRNA剪接变体。另一个目标 这些实验是生成表征蛋白质的抗体 急性炎症期间E-选择素的表达。时间课程和 将研究浸润白细胞的表型 免疫细胞化学和这些数据与E-的时间过程相比 选择素基因表达。 该项目的目的是获得洞察力 进入调节E-表达的细胞和分子机制 selectin。长期目标包括应用这些信息 急性和慢性炎症研究的调节机制 泌尿生殖道的疾病。据我所知 实验室拥有研究这种新方面的分子工具 E-选择素，体内。从这些拟议的研究中获得的数据将 对影响泌尿生殖器官的许多疾病的影响，并设置 以后调查的阶段着重于慢性炎症性疾病 （即间质性膀胱炎，前列腺炎，炎症方面 前列腺肥大）。