CRP, Diabetes, Atherothrombosis

在先前的提案中，中心假设是确定CRP是否通过对两者的影响来促进动脉粥样硬化血栓形成 内皮细胞和单核细胞。我们现在已经实现了这项提案的所有四个目标，并推动了这一领域的发展 关于C反应蛋白的血管效应。综上所述，我们阐明了C反应蛋白的分子机制 抑制eNOS(体外和体内)，我们已经记录了Fc-γ受体在以下生物学效应中的作用： C反应蛋白对内皮细胞、巨噬细胞和Wistar大鼠的影响。此外，我们还阐明了CRP诱导的机制。 剪切力下的单核细胞黏附，最后我们在Wistar大鼠体内证实了CRP具有 促进动脉粥样硬化的作用包括刺激NADPH-氧化酶、超氧化物歧化、MPO释放、氧化 巨噬细胞摄取低密度脂蛋白、组织因子、释放基质金属蛋白酶-9，降低血管反应性。糖尿病是促炎性疾病 以高C反应蛋白水平为特征的状态。然而，目前研究这一问题的数据很少。 C反应蛋白在促进糖尿病患者促炎状态中的作用。我们已经在令人兴奋和新颖的初步数据中表明 C反应蛋白在体内加重糖尿病环境(自发性糖尿病BB)中的促炎、促氧化作用 鼠)。因此，在这个相互竞争的更新中，我们希望进一步探讨CRP在糖尿病和动脉粥样硬化血栓形成中的作用。 为此，我们提出了两个具体目标。在具体目标1中，我们将继续扩大我们令人兴奋的 初步研究发现，CRP可加重糖尿病BB大鼠的促炎、促氧化状态。在这 模型，我们将确认CRP是否在体内加剧了糖尿病环境中的促炎和促氧化作用 阐明了C反应蛋白通过体内小干扰RNA和小干扰RNA发挥上述作用的分子机制(S) 反义寡核苷酸对不同的通路进行鉴定。基于主要来自我们团队和其他人的发现， 在特定目标2中，使用自发性糖尿病BB大鼠，CRP促进促凝血表型，我们将 现在在体内测试CRP对糖尿病环境中血栓形成的影响。此外，我们还将通过以下方式来阐明机制(S) 在糖尿病状态下，C反应蛋白促进动脉粥样硬化血栓形成。我们相信这些研究将提供更多新颖的数据。 支持C反应蛋白促进体内动脉粥样硬化血栓形成和促凝剂、促炎的假设 糖尿病的表型。探讨C反应蛋白增强氧化应激和细胞毒作用的分子机制 糖尿病环境中的炎症最终将导致针对减少炎症和氧化的治疗 糖尿病的应激反应和导致血管病变的减少

In the previous proposal, the central hypothesis was to determine if CRP promotes atherothrombosis by effects on both endothelial cells and monocytes. We have now executed all four aims of this proposal and have advanced the field with regards to the vascular effects of CRP. In summary, we have elucidated the molecular mechanism by which CRP inhibits eNOS (in-vitro and in-vivo), we have documented the role of Fc-gamma receptors in the biological effects of CRP on endothelial cells, macrophages and in Wistar rats. Furthermore, we have elucidated the mechanism of CRPinduced monocyte adhesion under shear stress, and finally we have confirmed in-vivo, in Wistar rats, that CRP has effects that promote atherosclerosis including stimulation of NADPH-oxidase, superoxide, MPO release, oxidized LDL uptake, tissue factor, MMP-9 release from macrophages and decreased vasoreactivity. Diabetes is a proinflammatory state that is characterized by high CRP levels. However, there is a paucity of data examining the role of CRP in promoting the pro-inflammatory state in diabetes. We have shown in exciting and novel preliminary data that CRP exacerbates in-vivo the pro-inflammatory, pro-oxidant effects in the diabetic milieu (spontaneously diabetic BB rat). Thus, in this competing renewal, we wish to further explore the effects of CRP on diabetes and atherothrombosis. To this end, we are proposing two specific aims. In specific aim 1, we will continue to expand our exciting preliminary findings that CRP accentuates the pro-inflammatory, pro-oxidant state in the diabetic BB rat. In this model, we will confirm if CRP exacerbates in-vivo the pro-inflammatory, pro-oxidant effects in the diabetic milieu and also elucidate the molecular mechanism (s) by which CRP exerts these effects by employing in-vivo siRNA and antisense oligonucleotides to the different pathways identified. Based on findings largely from our group and others, that CRP promotes a pro-coagulant phenotype, in Specific Aim 2, using the spontaneously diabetic BB rat, we will now test in-vivo the effect of CRP on thrombosis in the diabetic milieu. Also, we will elucidate the mechanism (s) by which CRP promotes atherothrombosis in the diabetic state. We believe these studies will provide further novel data in support of the hypothesis that CRP promotes atherothrombosis in-vivo and a procoagulant, pro-inflammatory phenotype in diabetes. Probing into the molecular mechanisms by which CRP augments oxidative stress and inflammation in the diabetic milieu will eventually lead to therapies targeted at reducing inflammation and oxidative stress in diabetes and resulting in a decrease in vasculopathies