Formation of Omega 3-Derived Electrophiles During Inflammation

描述（由申请人提供）：动脉粥样硬化占所有心血管疾病死亡人数的四分之三，预计到2020年动脉粥样硬化血栓性疾病将成为全球死亡的主要原因，是一个主要的健康负担。Omega-3 （w-3）多不饱和脂肪酸具有有效的抗炎作用和有益心血管的作用。虽然这些特性已被广泛认识，但其潜在机制仍不甚清楚。巨噬细胞活化过程中形成的脂质亲电图谱显示，细胞内存在酮衍生的w-3脂肪酸- omega -3脂肪酸，浓度高达200 nM。这些物种的形成依赖于COX-2，服用阿司匹林增加了这些w-3亲电脂肪酸氧衍生物（EFOX）的水平。最近，EFOX已被证明在人类巨噬细胞、中性粒细胞和动物炎症组织中形成。此外，EFOX已被发现形成谷胱甘肽和蛋白质加合物，并通过激活Nrf2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。该提案的假设是，EFOX是炎症和动脉粥样硬化过程中形成的具有抗动脉粥样硬化特性的生物学相关分子，通过信号通路的亲电调节转导w-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（即氘化和标记标准），以帮助严格的生物鉴定和定量的EFOX。这些标准将用于分析巨噬细胞的生物合成途径，并评估相关小鼠动脉粥样硬化模型的体内形成。这些物种及其靶点的生化和信号特性将使用细胞模型进行分析。最后，将在小鼠动脉粥样硬化模型上测试EFOX的药理作用。在完成时，我们期望确定不同的EFOX异构体物种及其在体内的相关性和形成。此外，我们希望能够清楚地了解它们的亲电反应性、2期基因表达的调节，特别是Nrf2/KEAP1通路和它们的PPARg激活。期望这一建议将有助于更好地了解w-3脂肪酸的作用，并改进减少动脉粥样硬化及其有害影响的药理学方法。

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 (w-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 w-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 w-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 w-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 PPARg activation. It is expected that this proposal will lead to a better understanding of w-3 fatty acid effects and to improved pharmacological approaches to decrease atherosclerosis and its detrimental effects.