Glutathionylated Products of Radical-Induced Lipid Oxidation in Inflammatory Disease

对FREE产生的产品的结构和生物活性的有限知识 自由基诱导的视网膜脂质氧化是临床进展的障碍。 有用的新疾病标记物和机械知情的治疗干预。明白-- ING细胞对自由基诱导的脂质氧化产物的反应是复杂的，因为它们 多样性和模拟酶产生的受体激动剂的能力。他们未被认可的 几乎可以肯定的是，世代和生物活动导致了目前的平庸疗效 可用于老年性黄斑变性(AMD)的治疗措施。我们发现 由氧化截短的花生四烯基磷脂产生的谷胱甘肽(GSH)衍生物 是半胱氨基(Cys)白三烯(LTS)的结构和功能类似物。我们把它们称为 伪(ø)LTS。我们将检验这些假设，这些假说认为有氧运动对视网膜病理和 光致视网膜变性大鼠模型的生理学研究。初步研究表明，ólts是 存在于人类视网膜中，在体内因氧化损伤而产生，并表现出类似LT的特征 生物活动。因此，LTS可以引起细胞反应，错误地认为是 仅由CysLts诱导。因此，它们是口译中的一个混杂因素 LTS参与视网膜病理生理的研究进展。建议数 研究将检验以下假设：LTS和相关的GSH衍生物被氧化截断 二十二碳六烯酰磷脂有助于受体依赖的炎症细胞因子信号- ING、视网膜水肿、病理性新生血管或重要的生理性视网膜病变 色素上皮(RPE)自噬。它们潜在的病理生理学意义 脂质氧化的谷胱甘肽产物在于它们在 高水平，例如，与CysLts的水平相比。我们将检验初步研究的结论 LTs是LT受体激动剂，并确定它们是否有能力诱导IL-13的表达 转化生长因子-β1促进LT的生物合成。因为交叉反应可能会混淆解释 在免疫分析中，我们用LC-MS/MS对LTS和LTS进行定量，我们将确定信号通路- 方法，例如，通过RNA-Seq研究，由自由基诱导的谷胱甘肽基化产物激活 脂质氧化解决问题：它们是否促进了眼部病理中的炎症 或者启动RPE细胞的自噬，它们是如何做到这一点的？分子水平上对角色的洞察 氧化应激在AMD等炎症性眼病发病机制中的作用 治疗干预措施的发展，以改善这些常见但 致残性疾病。

Limited knowledge of the structures and biological activities of products generated by free radical-induced lipid oxidation in the retina is a barrier to progress in the development of clinically useful new disease markers and mechanistically informed therapeutic interventions. Understand- ing cellular responses to free radical-induced lipid oxidation products is complex owing to their diversity and ability to mimic enzymatically generated receptor agonists. Their unrecognized generation and biological activities almost certainly contribute to the mediocre efficacy of currently available therapeutic measures for age related macular degeneration (AMD). We discovered glutathione (GSH) derivatives produced from oxidatively truncated arachidonyl phospholipids that are structural and functional analogues of cysteinyl (Cys) leukotrienes (LTs). We refer to them as pseudo (ø)LTs. We will test the hypotheses that øLTs contribute to retinal pathology and physiology in a rat model of light-induced retinal degeneration. Pilot studies show that øLTs are present in human retina, are produced in vivo consequent to oxidative insult, and exhibit LT-like biological activities. Therefore, øLTs can elicit cellular responses erroneously presumed to be induced exclusively by CysLTs. Consequently, they are a confounding factor for interpreting previous studies on the involvements of LTs in retinal pathology and physiology. The proposed research will test the hypotheses that øLTs and related GSH derivatives of oxidatively truncated docosahexaenoyl phospholipids contribute to receptor-dependent inflammatory cytokine signal- ing, retinal edema, pathological neovascularization, or physiologically important initiation of retinal pigment epithelium (RPE) autophagy. The potential pathophysiological significance of these glutathionylated products of lipid oxidation lies in the possibility of their ubiquitous generation in high levels, e.g., compared to those of CysLTs. We will test the conclusion of preliminary studies that øLTs are LT receptor agonists and determine if their ability to induce the expression of IL-13 and TGF-b1 promotes LT biosynthesis. Because cross reactivity may confound the interpretation of immunoassays, we quantitate øLTs and LTs by LC-MS/MS. We will determine signaling path- ways, e.g., with RNA-Seq studies, activated by glutathionylated products of free radical-induced lipid oxidation addressing the questions: do they promote inflammation in ocular pathology or initiate autophagy in RPE cells and how do they do it? Molecular level insights into the role of oxidative stress in the pathogenesis of inflammatory eye diseases such as AMD can facilitate development of therapeutic interventions that ameliorate the progression of these common but disabling diseases.