Formation of Omega 3-Derived Electrophiles During Inflammation

描述（由申请人提供）：动脉粥样硬化占心血管疾病所有死亡的四分之三，预计动脉粥样硬化血栓形成疾病将成为2020年全球主要死亡原因，是一个主要的健康负担。欧米茄-3（ω-3）多不饱和脂肪酸具有有效的抗炎作用和有益的心血管作用。虽然这些特性被广泛认可，但其潜在机制在很大程度上仍然未知。在巨噬细胞活化期间形成的脂质的亲电映射显示酮衍生的ω-3脂肪酸ω 3衍生的脂肪酸以高达200 nM的细胞内浓度存在。这些物质的形成依赖于考克斯-2，并且给予阿司匹林增加了这些ω-3亲电脂肪酸氧代衍生物（EFOX）的水平。最近，EFOX已被证明是在人类巨噬细胞，嗜中性粒细胞和动物组织受到炎症形成。此外，已发现EFOX形成谷胱甘肽和蛋白质加合物，并通过激活Nrf 2途径、抑制细胞因子表达和降低诱导型一氧化氮合酶水平来调节炎症途径。该建议的假设是EFOX是在炎症和动脉粥样硬化期间形成的具有抗动脉粥样硬化性质的生物学相关分子，其通过信号通路的亲电调节来抑制ω-3脂肪酸和阿司匹林的有益作用。为了解决这一假设，我们建议化学生成分析工具（即氘代和标记的标准品），以帮助严格的生物学鉴定和定量EFOX。这些标准品将用于分析巨噬细胞中的生物合成途径，并评估动脉粥样硬化相关鼠模型中的体内形成。将使用细胞模型分析这些物种及其靶标的生化和信号传导特性。最后，将在小鼠动脉粥样硬化模型上测试EFOX的药理学作用。在完成时，我们预计已经确定了不同的EFOX异构体种类及其体内相关性和形成。此外，我们希望能够清楚地了解它们的亲电反应性，2相基因表达的调节，特别是Nrf 2/KEAP 1通路及其PPARγ活化。预计这一建议将导致更好地理解ω-3脂肪酸的作用，并改进减少动脉粥样硬化及其有害作用的药理学方法。

DESCRIPTION (provided by applicant): Atherosclerosis accounts for three fourth of all deaths from cardiovascular disease and atherothrombotic diseases are projected to be the leading cause of death worldwide in 2020, representing a major health burden. Omega-3 (ω-3) polyunsaturated fatty acids exert potent anti-inflammatory actions and beneficial cardiovascular effects. Although these properties are widely recognized, the underlying mechanisms remain largely unknown. An electrophilic mapping of lipids formed during macrophage activation showed that keto-derived ω-3 fatty acids omega 3 derived-fatty acids were present at intracellular concentrations up to 200 nM. The formation of these species was dependent on COX-2, and administered aspirin increased the levels of these ω-3 electrophilic fatty acid oxo-derivatives (EFOX). Recently, EFOX have been shown to be formed in human macrophages, neutrophils and animal tissues subjected to inflammation. Additionally, EFOX have been found to form glutathione and protein adducts and to modulate the inflammatory pathways through activation of Nrf2 pathway, inhibition of cytokine expression and reduction of inducible nitric oxide synthase levels. The hypothesis of this proposal is that EFOX are biologically relevant molecules with anti-atherogenic properties formed during inflammation and atherosclerosis that transduce the beneficial effects of ω-3 fatty acids and aspirin through the electrophilic regulation of signaling pathways. To address this hypothesis we propose to chemically generate analytical tools (i.e. deuterated and labeled standards) to aid in the rigorous biological identification and quantitation of EFOX. These standards will be used for the analysis of biosynthetic pathways in macrophages and to assess the in vivo formation in a relevant murine model of atherosclerosis. The biochemical and signaling properties of these species and its targets will be analyzed using cellular models. Finally, the pharmacological effects of EFOX will be tested on a murine atherosclerosis model. At completion, we expect to have determined the different EFOX isomeric species and their in vivo relevance and formation. In addition, we expect to have gained a clear understanding of their electrophilic reactivities, modulation of phase 2 gene expression, in particular the Nrf2/KEAP1 pathway and their PPARγ activation. It is expected that this proposal will lead to a better understanding of ω-3 fatty acid effects and to improved pharmacological approaches to decrease atherosclerosis and its detrimental effects.