Cellular Reductive State Regulates Arteriogenesis

项目摘要 由进行性血管闭塞引起的组织缺血的发生率正在上升，并导致 几种以动脉阻塞为特征的心血管疾病，如外周动脉疾病。 组织血运重建对时间敏感，对于恢复足够的血流至关重要。减少 抗氧化能力，如还原型谷胱甘肽（GSH）浓度降低， 氧化应激的相应增加是疾病进展的标志， 功能障碍谷胱甘肽的减少被认为与疾病严重程度的线性增加相对应， 是一种不为人知的关系目前的建议旨在：（a）确定改变的影响 GSH：GSSG水平对蛋白谷胱甘肽化驱动血管内皮生长因子受体2（VEGFR 2）的影响 （B）确定谷胱甘肽化在氧化和剪切应力诱导的血管生成中的作用， 内皮细胞NF-κB信号传导，（c）在具有内皮细胞NF-κ B信号传导中的突变的鼠模型中研究体内动脉生成， 谷胱甘肽合成途径，并正在进行结扎，以模拟急性和慢性外周动脉疾病， 和（d）通过刺激更还原的细胞环境来恢复有缺陷的动脉生成进展， 改善内皮细胞功能。我们将检验中心假设，即 还原性和氧化性细胞环境驱动最佳VEGFR 2信号传导以介导动脉生成 重塑响应于增加的剪切和氧化应激。拟议的目标将利用体外 从我们的谷胱甘肽合成突变小鼠动物中分离的内皮细胞培养物， 专注于驱动VEGFR 2特异性信号传导的蛋白质的谷胱甘肽化。拟议的目标还包括 我们的动脉阻塞的体内小鼠模型作为血管重塑的临床相关模型。具体 目的1将着重于确定谷胱甘肽化在内皮细胞增殖过程中VEGFR 2活化中的作用。 动脉信号具体目标2将评估低水平氧化应激的作用及其对 谷胱甘肽化驱动动脉信号。我们将利用体外培养的内皮细胞分离， 我们的谷胱甘肽合成突变鼠动物研究目标1和2中的信号传导。具体目标3将评估 氧化/还原平衡在体内动脉生成重塑中的作用。在这里，我们将使用我们的体内 小鼠动脉阻塞模型。 该项目的成功完成将为谷胱甘肽的作用机制提供新的见解。 调节动脉结扎后GSH：GSSG生理范围内的动脉生成。此类信息可 为开发新的介入疗法以精确控制动脉后动脉生成奠定基础 堵塞。通过操纵谷胱甘肽和谷胱甘肽增强组织的血管重塑潜力 蛋白质谷胱甘肽化可能代表了减轻由于血管紧张素Ⅱ引起的组织损伤的关键的第一步 闭塞

PROJECT ABSTRACT The incidence of tissue ischemia resulting from progressive vascular occlusion is on the rise, and leads to several cardiovascular pathologies characterized by arterial blockage such as peripheral artery disease. Revascularization of tissue is time sensitive and essential to restore adequate blood flow. Decreases in antioxidant capacity such as decreases in the reduced form of glutathione (GSH) concentrations and corresponding increases in oxidant stress are hallmarks of disease progression and endothelial cell dysfunction. Decreases in glutathione are thought to correspond with a linear increase in disease severity that is a poorly understood relationship. The current proposal seeks to: (a) determine the influence of changing GSH:GSSG levels on protein glutathionylation driving vascular endothelial growth factor receptor 2 (VEGFR2) signaling in arteriogenesis, (b) determine the role of glutathionylation in oxidative and shear stress induced endothelial cell NF-κB signaling, (c) study in vivo arteriogenesis in murine models that have mutations in the GSH synthesis pathway, and are undergoing ligations to mimic acute and chronic peripheral artery disease, and (d) restore defective arteriogenesis progression by stimulating a more reductive cellular environment to improve endothelial cell function. We will test the central hypothesis that a critical balance between the reductive and oxidative cellular environments drives optimal VEGFR2 signaling to mediate arteriogenic remodeling in response to increased shear and oxidant stress. The proposed aims will utilize in vitro cultures of endothelial cells isolated from our glutathione synthesis mutant murine animals to generate data focusing on glutathionylation of proteins driving VEGFR2 specific signaling. The proposed aims also include our in vivo mouse models of arterial blockage as clinically relevant models of vascular remodeling. Specific Aim 1 will focus on determining the role of glutathionylation in VEGFR2 activation during endothelial cell arteriogenic signaling. Specific Aim 2 will assess the role of low level oxidant stress and its control over glutathionylation driving arteriogenic signaling. We will utilize in vitro cultures of endothelial cells isolated from our glutathione synthesis mutant murine animals to study signaling in aims 1 and 2. Specific Aim 3 will assess the role of the oxidative/reductive balance in arteriogenesis remodeling in vivo. Here we will use our in vivo mouse models of arterial blockage. Successful completion of this project will provide new insights into the mechanism by which glutathione regulates arteriogenesis in a physiologic range of GSH:GSSG following arterial ligation. Such information could be the basis for new intervention therapies developed to precisely control arteriogenesis following artery blockage. Enhancing the vascular remodeling potential of tissue through manipulation of glutathione and protein glutathionylation may represent a critical first step in attenuating tissue damage due to vascular occlusion.