Complement and Ischemic Acute Renal Failure

描述(由申请人提供)： 缺血性急性肾功能衰竭在医院环境中仍然是肾功能衰竭的常见原因。尽管进行了大量的研究，但这种类型的肾功能衰竭的预后并没有明显改善。我们和其他人已经证明，补体激活在小鼠缺血性急性肾功能衰竭的发展过程中是一个重要事件。本研究旨在分析缺血性急性肾功能衰竭模型中补体激活和补体依赖性损伤的机制。还将进行实验，以确定新产生的替代补体途径的抑制剂是否在这些损伤模型中具有治疗效果。 鉴于肾脏缺血损伤的复杂性，将使用几种体内和体外模型。这些包括在体内诱导小鼠缺血性急性肾功能衰竭，以及在存在或不存在补体因子的情况下使新鲜分离的肾小管受到缺氧的影响。许多体内实验将利用缺乏特定补体激活途径成分或补体抑制物的小鼠(即基因敲除小鼠)。用于测量这些实验终点的技术包括血清尿素氮分析、光学显微镜、培养细胞和组织切片的免疫荧光分析、ELISA法、西方印迹分析、基因阵列分析和定量RT-PC1L。此外，我们还将使用一些由我们的合作者开发的独特的分析方法，例如测量caspase活性的方法。 这些实验旨在深入分析肾缺血/再灌流过程中补体依赖的损伤机制。所使用的技术将利用补体生物学和急性肾功能衰竭领域的专家的专业知识。这些研究的目的是通过阻断这些致病事件来改善缺血性ARF，并开发降低与人类缺血性ARF相关的发病率和死亡率的策略。

DESCRIPTION (provided by applicant): Ischemic acute renal failure remains a common cause of renal failure in the hospital setting. In spite of substantial investigation, the prognosis for this type of renal failure has not significantly improved. We and others have shown that complement activation is an important event in the development of ischemic acute renal failure in mice. The proposed studies aim to analyze the mechanisms of complement activation and complement dependent injury in models of ischemic acute renal failure. Experiments will also be conducted to determine whether a newly created inhibitor of the alternative complement pathway is of therapeutic benefit in these models of injury. Given the complexity of ischemic injury in the kidney, several in vivo and in vitro models will be used. These include the in vivo induction of ischemic acute renal failure in mice, and subjection of freshly isolated renal tubules to hypoxia in the presence or absence of complement factors. Many of the in vivo experiments will utilize mice that are deficient in specific complement activation pathway components or in the inhibitors of complement (i.e. knockout mice). The techniques used to measure the endpoints of these experiments include assays for serum urea nitrogen, light microscopy, immunofluorescence of cells in culture and of tissue sections, ELISA, Westem blot analysis, gene array analysis, and quantitative RT-PC1L In addition, some unique assays developed by our collaborators will be utilized, such as a method of measuring caspase activation. These experiments are designed to analyze in depth the complement dependent mechanisms of injury which occur during renal ischemia/reperfusion. The techniques used will utilize the expertise of experts in the fields of complement biology as well as acute renal failure. The purpose of these studies is to ameliorate ischemic ARF by blocking these pathogenic events, and to develop strategies for decreasing the morbidity and mortality associated with ischemic ARF in humans.