MECHANISMS OF ISCHEMIC INJURY AND REPAIR

描述（由申请方提供）：拟定研究的总体目标 缺血性肾细胞损伤和修复的机制有待进一步研究 长期目标是建立一种有效的治疗方法， 治疗人类急性肾衰竭（ARF）。在上一个支持期， 已经发展并验证了缺血性ARF是一种疾病的假设， 白细胞-内皮细胞相互作用，特别是在外髓质， 肾脏，导致小血管充血和氧气受损 递送导致肾小管细胞损伤。受损的肾小管被激活 内皮细胞通过产生 细胞因子和趋化因子上调粘附受体， 白细胞我们最近建立了三种小鼠模型，其中肾脏 可以防止缺血性损伤。这些模型将被用来 评估假设并确定重要的候选蛋白质 造成伤害的因素或内源性保护过程。具体目标1 我们将使用缺血性损伤的保护模型来定义 外髓白细胞和内皮细胞活化的重要性， 缺血性急性肾衰竭的病理生理学相互作用。国 将评估S3节段的分化和应激反应， 考虑到改变的潜在重要性， 白细胞-内皮细胞相互作用相对于 近曲小管上皮细胞损伤。粘附分子和趋化因子 将研究参与以及肾损伤的潜在作用 白细胞-上皮相互作用中的分子-1（KIM-I）。具体目标2 将评估nmb对白细胞功能的作用， 与炎症有关的候选基因可能对缺血性心脏病 肾脏外髓质的损伤或诱导性保护过程。我们 将决定NMB及其三个潜在功能域的作用 (the多囊肾[PKD]结构域，以及YXXphi和双亮氨酸结构域， 胞质尾）对白细胞与内皮细胞结合的影响，以及对 吞噬作用将通过转录本进行基因表达分析。 分析与基因数据库查询相结合。我们将找出那些基因， 在三种情况下， 被鉴定为导致保护肾脏免受缺血性损伤。 免疫细胞化学、原位杂交、激光捕获显微切割和 TaqMan真实的时间RT-PCR将用于鉴定基因的亚组，其 表达模式在缺血后外周血中有差异调节， 当比较受保护的肾脏与未受保护的肾脏时，

DESCRIPTION (provided by applicant): The overall goal of the proposed studies remains to understand the mechanisms of ischemic kidney cell injury and repair with a long range goal to establish therapies that will be useful to prevent and treat acute renal failure (ARF) in man. Over the last support period we have developed and tested the hypothesis that ischemic ARF is a disease in which leukocyte-endothelial interactions, particularly in the outer medulla of the kidney, lead to congestion of the small vessels and impaired oxygen delivery with resultant tubule cell injury. The damaged tubule and activated endothelium contributes to additional vascular compromise by producing cytokines and chemokines which up regulate adhesion receptors and attract leukocytes. We have recently established three mouse models in which the kidney is protected against ischemic injury. These models will be exploited to evaluate the hypothesis and identify proteins that are candidates as important contributors to injury or endogenous processes of protection. In Specific Aim 1 we will use models of protection against ischemic injury to define the importance of outer medullary leukocyte and endothelial activation and interactions for the pathophysiology of ischemic acute renal failure. The state of differentiation and stress response of the S3 segment will be evaluated to put into context the potential importance of alterations in leukocyte-endothelial interactions relative to change in susceptibility to injury of the proximal tubule epithelial cell. Adhesion molecule and chemokine involvement will be studied as will the potential role of Kidney Injury Molecule-1 (KIM-I) in leukocyte-epithelial interactions. In Specific Aim 2 we will evaluate the role of nmb on leukocyte function and identify additional candidate genes implicated in inflammation that may be important for ischemic injury or inducible protective processes in the outer medulla of the kidney. We will determine the effect of nmb and three of its potential functional domains (the polycystic kidney [PKD] domain, and the YXXphi and di-leucine domains in the cytosolic tail) on binding of leukocytes to endothelial cells and on phagocytosis. Gene expression analyses will be performed by transcript profiling coupled to a gene database query. We will identify those genes that show coordinate up or down regulation under three conditions that we have identified to result in protection of the kidney against ischemic injury. Immunocytochemistry, in situ hybridization, laser capture microdissection and TaqMan real time RT-PCR will be used to identify that subgroup of genes whose expression pattern is differentially regulated in the postischemic outer medulla when comparing protected vs non-protected kidneys.