Ethyl Pyruvate: A Novel Treatment for Sepsis

丙酮酸乙酯：脓毒症的新型治疗方法

• 批准号: 6765286
• 负责人: Mitchell P. Fink
• 金额: $ 28.71万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6765286
• 关键词: alkylation; antioxidants; apoptosis; biological signal transduction; blood /lymphatic pharmacology; blood disorder chemotherapy; enzyme linked immunosorbent assay; ethanol; free radical oxygen; gel mobility shift assay; glutathione; immunocytochemistry; inflammation; laboratory mouse; mitogen activated protein kinase; nuclear factor kappa beta; oxidation reduction reaction; pyruvates; septicemia; thiols; tissue /cell culture; western blottings; alkylation; antioxidants; apoptosis; biological signal transduction; blood /lymphatic pharmacology; blood disorder chemotherapy; enzyme linked immunosorbent assay; ethanol; free radical oxygen; gel mobility shift assay; glutathione; immunocytochemistry; inflammation; laboratory mouse; mitogen activated protein kinase; nuclear factor kappa beta; oxidation reduction reaction; pyruvates; septicemia; thiols; tissue /cell culture; western blottings

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.

描述（由申请人提供）： 丙酮酸乙基（EP）是由丙酮酸和乙醇形成的酯。在初步研究中，我们已经证明，EP可以改善肠道和肝损伤或改善生存率，当它用作治疗患有肠系膜缺血的啮齿动物和再灌注，出血性休克，内毒素性毒素，内毒素或多种细菌细菌细菌性sepsis时，它可以改善生存率。此外，我们证明了EP是对活性氧（ROS）的有效清除剂，并且我们已经表明，该化合物也是一种抗炎剂，可抑制促炎性信号传导因子的激活，NF-kappab和p38有丝分裂原激活的蛋白激活蛋白激酶。 EP还抑制了新型细胞因子的释放，高迁移率组框1（HMGB1）。在这些令人兴奋的观察结果的提示下，我们建议进行一系列实验，旨在更好地阐明负责EP的抗炎和治疗作用的机制。这项工作将以三个具体目标进行组织。目标1是检验三个可能解释EP的有益影响的广泛假设。这些假设是：ROS假设，烷基化假设和谷胱甘肽（GSH）耗尽假设。 ROS假设提出，EP充当抗氧化剂的能力解释了其细胞保护作用和抗炎作用。烷基化假设提出，EP充当烷基化的烷基硫醇基团，从而使重要的信号传导或效应分子失活，例如转录因子的亚基，NF-kappab的亚基或涉及凋亡过程的各种caspase。 GSH耗竭假说是烷基化假设的一种变体，提出EP烷基化GSH。 GSH的耗竭以有利于与NF-kappab亚基形成混合二硫化物的方式移动细胞氧化还原平衡，从而通过该途径干扰信号。目标2是对EP对NF-kappab激活的影响进行更详细的研究。 AIM 3是研究负责通过EP抑制HMGB 1释放的机制。所有这些目标将使用分子和药理学方法的组合进行，研究将采用体外（细胞培养）和体内（动物模型）方法。对EP的抗炎和治疗作用的机制有了更好的了解，可以识别与先天免疫反应有关的新型细胞途径。