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

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）： 丙酮酸乙酯（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，当 EP 用作治疗剂来治疗患有肠系膜缺血和再灌注、失血性休克、内毒素血症或多微生物细菌败血症的啮齿动物时，可以改善肠道和肝脏损伤或提高生存率。此外，我们还证明 EP 是活性氧 (ROS) 的有效清除剂，并且该化合物也是一种抗炎剂，可抑制促炎信号因子 NF-KappaB 和 p38 的激活有丝分裂原激活蛋白激酶。 EP 还抑制一种新型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，旨在更好地阐明 EP 抗炎和治疗作用的机制。这项工作将根据三个具体目标进行组织。目标 1 是测试可能解释 EP 有益效果的三个广泛假设。这些假设是：ROS 假设、烷基化假设和谷胱甘肽 (GSH) 消耗假设。 ROS 假说提出 EP 作为抗氧化剂的能力解释了其细胞保护和抗炎作用。烷基化假说提出 EP 作为亲电子试剂发挥作用，将关键的硫醇基团烷基化，从而使重要的信号分子或效应分子失活，例如转录因子、NF-KappaB 的亚基或参与细胞凋亡过程的各种半胱天冬酶。 GSH 耗尽假说是烷基化假说的一个变体，提出 EP 使 GSH 烷基化。 GSH 的消耗会改变细胞氧化还原平衡，从而有利于与 NF-KappaB 亚基形成混合二硫化物，从而干扰通过该途径的信号传导。目标 2 是对 EP 对 NF-KappaB 激活的影响进行更详细的研究。目标 3 是研究 EP 抑制 HMGB 1 释放的机制。所有这些目标将结合分子和药理学方法来实现，并且研究将采用体外（细胞培养）和体内（动物模型）方法。更好地了解 EP 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细胞途径。