RESOLUTION OF DIABETIC VASCULAR INFLAMMATION: ROLE OF LIPID MEDIATORS PROJ5

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 项目5：消解糖尿病血管炎症：脂质介质的作用 该项目的总体目标是更好地了解支持和维持糖尿病慢性低度血管炎症的机制，以及是否可以刺激炎症的消退阶段来预防糖尿病的血管功能障碍。我们的假设是，糖尿病期间的慢性血管炎症可能部分归因于促进炎症消退的内源性逆调节脂质中介途径的缺失。因此，用合成的促进分解的脂质介质，如脂氧素和分解素，修复这些缺陷，可以促进与长期糖尿病相关的血管炎症的解决。为了验证这一新的假设，我们设计了以下具体目标： 1.观察糖尿病急性炎症消退的变化。为此，我们将在微生物腹膜炎的小鼠模型中测量白细胞渗透的强度和持续时间，在该模型中，我们定义了特定的分辨率指数。炎症的时间进程将在非糖尿病小鼠以及1型和2型糖尿病的小鼠模型中进行研究。1型糖尿病将通过注射链脲佐菌素建立，而为了研究2型糖尿病，将使用db/db小鼠。这些模型的比较将有助于评估高血糖和高胰岛素血症在糖尿病炎症消退中的作用，并有助于理解糖尿病影响炎症消退的机制； 2.评估糖尿病患者分解前期脂质介体生物合成改变对其分解能力改变的贡献。为了了解糖尿病是如何影响消退的，我们将使用目标液相色谱/质谱仪来确定腹膜炎期间脂类介体生物合成的变化。我们将确定在1型和2型糖尿病模型腹膜炎的时间过程中，哪些脂质介质途径(促炎性和促解性)受到影响。此外，为了阐明糖尿病影响前分解脂介体生物合成的机制，我们将测量它们的生物合成中间产物，并确定它们如何受到糖尿病的影响；以及 3.确定使用促进分解的脂质介体治疗是否能恢复糖尿病患者的分解改变。我们将确定外源性脂素和分解素治疗如何影响非糖尿病和糖尿病小鼠腹膜炎的缓解。为了阐明这些介质影响炎症的机制，我们将通过活体显微镜检查它们对白细胞的影响：内皮细胞的相互作用。为了阐明前分解脂介体对抗糖尿病血管炎症的细胞机制，我们将研究脂氧素和分解素如何影响高糖和高脂肪酸诱导的微血管内皮细胞信号通路。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Project 5: Resolution of diabetic vascular inflammation: Role of lipid mediators The overall aim of this project is to develop a better understanding of the mechanisms that support and sustain chronic low-grade vascular inflammation in diabetes and whether the resolution phase of inflammation could be stimulated to prevent vascular dysfunction in diabetes. Our hypothesis is that chronic vascular inflammation during diabetes could be attributed, in part, to a loss in endogenous counter-regulatory lipid mediator pathways that promote the resolution of inflammation. It follows that restoration of these deficits with synthetic pro-resolving lipid mediators, such as the lipoxins and resolvins, could facilitate the resolution of vascular inflammation associated with long-term diabetes. The following specific aims were designed to test this novel hypothesis: 1. Examine diabetic changes in the resolution of acute inflammation. For this, we will measure the intensity and duration of leukocyte infiltration in a murine model of microbial peritonitis in which we have defined specific resolution indices. The time course of inflammation will be studied in non-diabetic mice and in murine models of both type 1 and type 2 diabetes. Type 1 diabetes will be established by streptozotocin injections and for studying type 2 diabetes, db/db mice will be used. Comparisons between these models will help in assessing the contribution of hyperglycemia and hyperinsulinemia to diabetic changes in the resolution of inflammation and in understanding the mechanism by which diabetes affects the resolution of inflammation; 2. Assess the contribution of altered pro-resolution lipid mediator biosynthesis to diabetic changes in resolution. To understand how diabetes affects resolution, we will identify changes in lipid mediator biosynthesis during peritonitis using targeted liquid chromatography/mass spectrometry. We will determine which lipid mediator pathways (pro-inflammatory vs. pro-resolution) are affected during the time course of peritonitis in models of both type 1 and type 2 diabetes. Moreover, to elucidate the mechanisms by which diabetes affects pro-resolution lipid mediator biosynthesis, we will measure their biosynthetic intermediates and determine how they are affected by diabetes; and 3. Determine whether treatment with pro-resolution lipid mediators restores diabetic changes in resolution. We will determine how treatment with exogenous lipoxins and resolvins affects the resolution of peritonitis in non-diabetic and diabetic mice. To delineate the mechanisms by which these mediators affect inflammation, we will examine their effects on leukocyte:endothelial interactions by intravital microscopy. To elcuidate the cellular mechanisms by which pro-resolution lipid mediators counter-act diabetic vascular inflammation, we will examine how lipoxins and resolvins affect high glucose and high fatty acid-induced signaling pathways in microvascular endothelial cells.