Lipid mediators of inflammation and insulin resistance in adipocytes and adipose

描述（由申请人提供）：越来越多的证据支持亚急性炎症在胰岛素抵抗和2型糖尿病发病机制中的重要作用。流行病学研究将肥胖和胰岛素抵抗与广泛的脂肪源性炎症标志物相关联，并且生化和生理学研究表明脂肪组织源性细胞因子和趋化因子可充当介质。最近的发现还将脂肪组织（ATM）中的巨噬细胞与肥胖诱导的炎症的发展和胰岛素抵抗的发病机制联系起来。考虑到宿主防御中的急性炎症过程与肥胖中的慢性炎症之间的相似性，我曾问过我们是否可以利用丰富的急性炎症知识库来了解更多关于肥胖诱导的胰岛素抵抗的信息。促炎和抗炎脂质介质的产生是急性炎症的一个广泛领域，在肥胖研究中尚未完全研究和低估。多种内源性脂质促进（如类花生酸、洋地黄素）或帮助缓解（如消退素、脂氧素）急性炎症反应。我推测脂肪组织中的脂质介质也可能在肥胖诱导的胰岛素抵抗的发病机制中发挥作用。为了测试这一点，我计划分析完整脂肪组织和特别感兴趣的组成细胞类型中的生物活性脂质：脂肪细胞和两种ATM群体，即富脂（泡沫细胞）或贫脂（典型）。我们聘请了一位世界领先的脂质组学分析专家来协助我们，并帮助指导我的研究。我们计划使用的方法包括基于FACS的同质ATM亚型的纯化，以及基于气相色谱和液相色谱的质谱法（LC-UV MS/MS和GC-MS），再加上基于计算机的自动化系统，该系统配备了数据库和新的搜索算法，用于介体分析。一旦介质被确定，我将测试它们在培养的3 T3-L1脂肪细胞（例如胰岛素信号传导和胰岛素刺激的葡萄糖摄取）和培养的RAW细胞和分离的巨噬细胞（例如NF-κ B）上的活性。和靶基因激活）。 公共卫生部门：本提案中概述的研究提供了一种无偏倚的方法来识别可能参与胰岛素抵抗发展的脂质介质。我的方法不仅要确定潜在的介质，但也有新的机制，脂肪细胞和巨噬细胞之间的串扰，可能有助于肥胖诱导的胰岛素抵抗的发展。

DESCRIPTION (provided by applicant): Increasing evidence supports important roles for subacute inflammation in the pathogenesis of insulin resistance and type 2 diabetes. Epidemiological studies associate obesity and insulin resistance with a broad range of adipose-derived inflammatory markers, and biochemical and physiological studies suggest that adipose tissue-derived cytokines and chemokines may act as mediators. Recent discoveries also link macrophages in adipose tissue (ATMs) to the development of obesity-induced inflammation and the pathogenesis of insulin resistance. Given the similarities between acute inflammatory processes in host defense and the chronic inflammation seen in obesity, I have asked whether we might use the rich knowledge base for acute inflammation to learn more about obesity-induced insulin resistance. The generation of pro- and anti-inflammatory lipid mediators is a broad area of acute inflammation that has been incompletely studied and underappreciated in obesity research. A wide variety of endogenous lipids either promote (e.g. eicosanoids, prostaglandins) or help to resolve (e.g. resolvins, lipoxins) acute inflammatory responses. I hypothesize that lipid mediators in adipose tissue may also have roles in the pathogenesis of obesity-induced insulin resistance. To test this, I plan to analyze bioactive lipids in intact adipose tissue and component cell types of special interest: adipocytes and two populations of ATMs, those that are lipid-rich (foam cells) or lipid-poor (typical). We have recruited the assistance of a world leader in lipidomic analyses to assist us and help guide my studies. The methods we plan to use include FACS-based purification of homogenous ATM subtypes, and gas and liquid chromatography-based mass spectrometry (LC-UV MS/MS and GC-MS) coupled with a computer-based automated system equipped with databases and novel searching algorithms for mediator profiling. Once mediators are identified, I will test their activities on cultured 3T3-L1 adipocytes (e.g. insulin-signaling and insulin-stimulated glucose uptake) and both cultured RAW cells and isolated macrophages (e.g. NF-?¿ and target gene activation). PUBLIC HEALTH REVELANCE: The studies outlined in this proposal provide a non-biased approach to identifying lipid mediators potentially involved in the development of insulin resistance. My approach should not only identify potential mediators but also new mechanisms for cross-talk between adipocytes and macrophages that may contribute to the development of obesity-induced insulin resistance.