The Role of MIF in Endothelial Apoptosis

描述（由申请人提供）：内皮细胞（EC）损伤是败血症的主要特征。MIF是脓毒症期间循环和内皮细胞中升高的一种细胞因子，在脓毒症的病理生理中起着关键作用。我们实验室和其他人的研究表明，人类EC对凋亡刺激表现出高度的抗性，这与抗凋亡途径的上调一致。我们假设MIF在这些EC中起阻断细胞凋亡的作用。本研究的目的是通过以下目的来表征MIF在内皮细胞凋亡和活化中的作用：在SA #1中，我们将使用两种方法来表征MIF缺乏的人肺动脉内皮细胞（HPAEC）的凋亡：1)我们将使用RNA干扰技术下调HPAEC中MIF的产生；2)我们将破坏MIF受体与中和抗体的相互作用。然后将使用缺氧、化学/再灌注、TNF和内毒素治疗来测试表达mif和mif缺乏的HPAEC对凋亡刺激的敏感性。在SA#2中，我们定义了HPAEC对MIF刺激的已知和新的反应。最后，我们将描述SA#3中MIF缺失时暴露的凋亡通路。

DESCRIPTION (provided by applicant): Endothelial cell (EC) injury is a cardinal feature of sepsis. MIF, a cytokine elevated both in the circulation and endothelial cells during sepsis, plays a critical role in the pathophysiology of this disease. Studies in our lab and others demonstrate that human EC display a high resistance to apoptotic stimuli consistent with the upregulation of anti-apoptotic pathways. We hypothesize that MIF functions to block apoptosis in these EC. The goal of this study is to characterize the role of MIF in endothelial cell apoptosis and activation via the following aims: In SA #1 we will characterize apoptosis in human pulmonary artery endothelial cells (HPAEC) deficient in MIF using two approaches 1) we will downregulate MIF production in HPAEC using RNA interfering technology, and 2) we will disrupt MIF-receptor interactions with neutralize antibodies. The susceptibility of MIF-expressing and MIF-deficient HPAEC to apoptotic stimuli will then be tested using hypoxia, chemic/reperfusion, TNF, and endotoxin treatment. In SA#2 we define known and novel responses to MIF stimulation in HPAEC. Finally, we will characterize the apoptotic pathways exposed in the absence of MIF in SA#3.