Ethyl Pyruvate: A Novel Treatment for Sepsis

描述(由申请人提供)： 丙酮酸乙酯(EP)是丙酮酸和乙醇形成的酯。在初步研究中，我们已经证明，当EP被用作治疗肠系膜缺血再灌注、失血性休克、内毒素血症或多菌细菌败血症的啮齿动物时，它可以改善肠道和肝脏损伤或提高存活率。此外，我们还证明了EP是一种有效的ROS清除剂，我们还证明了该化合物也是一种抗炎剂，它抑制了促炎信号因子NF-kappaB和p38丝裂原活化蛋白激酶的激活。EP还抑制一种新的细胞因子--高迁移率族蛋白1(HMGB1)的释放。在这些令人兴奋的观察结果的推动下，我们建议进行一系列实验，以更好地阐明EP抗炎和治疗作用的机制(S)。这项工作将在三个具体目标下组织。目标1是测试三个可能解释EP有益影响的广泛假设。这些假说是：ROS假说、烷基化假说和谷胱甘肽(GSH)耗竭假说。ROS假说认为，EP作为抗氧化剂的能力解释了其细胞保护和抗炎作用。烷基化假说认为，EP作为一个电泳体，使关键的硫醇基团烷基化，从而使重要的信号或效应分子失活，如转录因子、核因子-kappaB的亚单位或参与细胞凋亡过程的各种caspase。GSH耗竭假说是烷基化假说的一个变种，它提出EP使GSH烷基化。谷胱甘肽的耗尽改变了细胞氧化还原平衡，有利于与核因子-kappaB亚单位形成混合二硫化物，从而干扰了这一途径的信号传递。目的2是对EP对核因子-kappaB活化的影响进行更详细的研究。目的3探讨EP抑制HMGB 1释放的机制。所有这些目标都将使用分子和药理学方法相结合的方法来实现，研究将使用体外(细胞培养)和体内(动物模型)方法。更好地了解EP的抗炎和治疗作用的机制可能有助于识别参与先天免疫反应的新的细胞通路。

DESCRIPTION (provided by applicant): Ethyl pyruvate (EP) is the ester formed from pyruvic acid and ethanol. In preliminary studies, we have documented that EP ameliorates intestinal and hepatic injury or improves survival when it is used as a therapeutic agent to treat rodents subjected to mesenteric ischemia and reperfusion, hemorrhagic shock, endotoxemia, or polymicrobial bacterial sepsis. In addition, we have demonstrated that EP is an effective scavenger of reactive oxygen species (ROS), and we have shown that this compound is also an anti-inflammatory agent that inhibits activation of the pro-inflammatory signaling factors, NF-KappaB and p38 mitogen-activated protein kinase. EP also inhibits release of a novel cytokine, high mobility group box 1 (HMGB1). Prompted by these exciting observations, we propose to carry out a series of experiments that are designed to better elucidate the mechanism(s) responsible for the anti-inflammatory and therapeutic effects of EP. The work will be organized under three Specific Aims. Aim 1 is to test three broad hypotheses that might account for the beneficial effects of EP. These hypotheses are: the ROS Hypothesis, the Alkylation Hypothesis, and the Glutathione (GSH) Depletion Hypothesis. The ROS Hypothesis proposes that EP's ability to function as an antioxidant accounts for its cytoprotective and anti-inflammatory effects. The Alkylation Hypothesis proposes that EP functions as an electrophile that alkylates key thiol groups and thereby inactivates important signaling or effector molecules, such as subunits of the transcription factor, NF-KappaB, or various caspases involved in the process of apoptosis. The GSH Depletion Hypothesis, a variant of the Alkylation Hypothesis, proposes that EP alkylates GSH. Depletion of GSH shifts the cellular redox balance in a way that favors the formation of mixed disulfides with NF-KappaB subunits and thereby interferes with signaling via this pathway. Aim 2 is to carry out more detailed studies on the effects of EP on NF-KappaB activation. Aim 3 is to investigate the mechanisms responsible for inhibition of HMGB 1 release by EP. All of these aims will be carried out using a combination of molecular and pharmacological approaches and the studies will employ both in vitro (cell culture) and in vivo (animal model) approaches. Achieving a better understanding of the mechanisms responsible for the anti-inflammatory and therapeutic effects of EP may permit identification of novel cellular pathways involved in the innate immune response.