Functional Char. of Pro-Inflammatory Pathways Influencing Insulin Influencing Ins

胰岛素抵抗是一种常见的代谢状态，它是X综合征(代谢综合征，在- 苏林耐药综合征)。X综合征与肥胖和2型糖尿病密切相关， 在美国持续流行的肥胖症使这一系列异常现象更加普遍。 这个项目的一个主要主题是慢性炎症在HU的病因中起着核心作用。 男性处于胰岛素抵抗状态。此外，我们认为巨噬细胞可能是这种炎症的始动细胞。 血管紧张素诱导的胰岛素抵抗。支持这一概念的有力数据已经来自最近的研究 在巨噬细胞敲除动物中，我们证明了在体内禁用炎症或途径 巨噬细胞导致小鼠的整体胰岛素敏感状态，而过度激活(或降压) 巨噬细胞内的炎症途径导致糖耐量异常、高胰岛素血症和 胰岛素抵抗。在这项提案中，我们计划进行一系列广泛的体外和体内研究 目的：明确巨噬细胞在炎症诱导的胰岛素抵抗中的全部作用，以及巨噬细胞在炎症诱导的胰岛素抵抗中的作用。 脱轨装置。我们将使用基因芯片的方法来阐明受PPARy调控的基因网络 以及STS-LI脂肪细胞中的两个共抑制因子N-CoRand和SMRT。我们已经开发出了体外检测方法， 使我们能够剖析巨噬细胞行程的各个步骤，包括内皮细胞的迁移， 趋化性和巨噬细胞与脂肪细胞的直接结合。此外，我们还建立了共同的文化 巨噬细胞的加入引起STS-LI脂肪细胞的细胞胰岛素抵抗的系统，以及一个数字- 提出了通过实验来确定这一效应的基本机制。最后，我们将大力 通过使用一些新的基因敲除模型和新策略来继续我们的小鼠研究。在本AP中- 方法，我们将大量使用给定的KO动物的骨髓移植到受辐射的动物 C57BL/6将在相对较高的吞吐范围内创建功能性巨噬细胞KO模型。我们也会这样做 利用我们的慢RNA载体治疗正常骨髓的实验研究 巨噬细胞内特定的炎症途径成分。这就产生了一种骨髓型的 然后，这些细胞可以被移植并植入受辐射的宿主小鼠体内。这 进一步提高了我们方法的吞吐量，并且还允许多路传输敲除 评估组合效应。因此，这些结果应该确定激活的相关机制 巨噬细胞导致胰岛素靶组织的全局性胰岛素抵抗，也应该更好地阐明 目前可用的TZD的作用机制和组织作用部位。

Insulin resistance is a common metabolic condition which underlies Syndrome X(Metabolic Syndrome, In- sulin Resistance Syndrome). Syndrome Xis strongly associated with obesity and Type 2 diabetes and the ongoing obesity epidemicin the United States makes this constellation ofabnormalities even more prevalent. An overarchingtheme in this project is that chronic inflammationplays a central role in the etiology of hu- man insulin resistant states. Further, wepropose that the macrophage canbe an initiator ofthis inflamma- tion-induced insulin resistance. Powerful data to support this concept has already come from recent studies of macrophageknockout animals in whichwe showed that disabling the inflammatorypathwaywithin macrophages leads to a state ofglobal insulin sensitivity in mice,whereas, hyperactivating (or derepressing) the inflammatory pathway within macrophages leads to a state ofglucose intolerance, hyperinsulinemia, and insulin resistance. In this proposal, we plan to conduct an extensive series ofin vitro and in vivo studies aimed at identifying the full role ofthe macrophagein inflammation-induced insulin resistance and the un- derlying mechanisms. We will use a gene array approachto elucidate the gene network regulated by PPARy and the two co-repressors N-CoRand SMRT in sTs-Li adipocytes. Wehave developed in vitro assays which allow us to dissect out the various steps ofthe macrophageitinerary, including endothelial transmigration, chemotaxis, and direct binding of macrophages to adipocytes. In addition, we have established a co-culture system in which addition ofmacrophages causes cellular insulin resistance in sTs-Li adipocytes, and a num- ber of experiments are proposed to identify the basic mechanisms ofthis effect. Finally, we willvigorously pursue our mouse studies by using a number ofnewknockout models as well as new strategies. In this ap- proach, we will make heavyuse ofbone marrowtransplantation from a given KOanimal into irradiated C57bl/6 hosts to create functional macrophage KO models on a relativelyhigh throughput scale. Wewill also utilize our lentisiRNAmethod oftreating normal bone marrow with lentisiRNAvectors targeted against spe- cific inflammatory pathway components within macrophages. This creates a bone marrowknockdownof the desired target, and these cells can then be transplanted and engraftedinto an irradiated host mouse. This further enhances the throughput of our approach and also allows the possibility of multiplexing knockouts to assess combinatorial effects. Assuch, these results should identify relevant mechanisms whereby activation of macrophages causes global insulin resistance in insulin target tissues and should also better elucidate the mechanisms of action, and tissue sites of action of currently available TZDs.