MECHANISMS OF VASCULAR REMODELING USING IN VITRO 3D

血管重塑是一个涉及内皮细胞的复杂过程 影响血管壁平滑肌细胞的损伤/功能障碍 增殖、肥大、迁移和代谢，是一个贡献者 与高血压、动脉粥样硬化等疾病的发病机制有关 血管再狭窄。然而，血管生成的细胞和分子机制 重塑还没有被很好地理解。我们提出了一项综合研究，使用 具有正常血管细胞的静态和血流动力学三维模型 和我们从盐敏感型高血压中分离到的那些 大鼠，模拟血管壁的结构并提供 它的物理环境。这种方法将允许解决 假设致动脉粥样硬化的低密度脂蛋白和低密度脂蛋白 血流动力包括血管中细胞和分子的变化。 通过诱导血管壁的机械转导机制来实现细胞 会导致血管重塑。我们将使用选定的端点 (形态表型、细胞形状和面积、细胞增殖和基因 表达)，已被报道为特有的血管细胞 在重塑过程中的变化。为了解决这一假设，我们 提出以下具体目标：1)评估形态变化 表型、细胞骨架排列、有丝分裂标记和通透性 经内皮层三维血管细胞共培养模型的建立 是否接受了高水平的低密度脂蛋白治疗；2)测量时间 早期反应基因(c-myc和c-fos)表达的变化， 生长因子(PDGF和转化生长因子/β)、细胞外基质蛋白(层粘连蛋白 和III型胶原)和NO合成酶 低密度脂蛋白；3)使用抑制剂，评估钙、蛋白激酶和 MAP-K在低密度脂蛋白作用早期信号机制中的作用 在血管细胞上使用这两个模型。拟议的研究将提供 低密度脂蛋白致动脉粥样硬化水平改变的机制 改良高血压的形态和功能表型 血管内皮细胞和平滑肌细胞。建议的长期目标是 研究是为了更好地理解时间序列 血管早期参与的事件(和信号) 改建。所获得的知识将为发展战略提供建议 治疗不同血管疾病的最佳疗法。

Vascular remodeling is a complex process involving endothelial cell injury/dysfunction which affects vascular wall smooth muscle cell proliferation, hypertrophy, migration and metabolism, and is a contributor to the pathogenesis of disorders such as hypertension, atherosclerosis and restenosis. However, the cellular and molecular mechanisms of vascular remodeling are not well understood. We propose an integrated study using static and hemodynamic 3-dimensional models with normal vascular cell lines and those that we have isolated from salt sensitive hypertensive rats, that mimics the architecture of the vascular wall and also provide its physical environment. This approach will allow to address the hypothesis that the combination of atherogenic levels of LDL and hemodynamic forces include cellular and molecular changes in vascular cells by inducing mechanotransduction mechanisms for the blood vessel wall that result in vascular remodeling. We will use selected end-points (morphological phenotype, cell shape and area, cell proliferation and gene expression), which have been reported as characteristic vascular cell changes during the remodeling process. To address this hypothesis, we propose the following specific aims: 1) Assess changes in morphological phenotype, cytoskeleton arrangement, mitogenic labeling and permeability through the endothelial layer when the 3-D vascular cell co-culture models are treated or not treated with high levels of LDL; 2) Measure temporal changes in the expression of early response genes (c-myc and c-fos), growth factors (PDGF and TGF/beta), extracellular matrix proteins (laminin and collagen III) and NO synthase following treatment of cultures with LDL; 3) Using inhibitors, assess the role of calcium, protein kinases and MAP-kinase in the early signaling mechanisms mediating the effects of LDL on vascular cells using the two models. The proposed studies will provide insight into the mechanism by which atherogenic levels of LDL alter the morphological and functional phenotypes of hypertension-modified endothelial and smooth muscle cells. The long term goal of the proposed studies is to provide a better understanding of the temporal sequence of events (and signaling) involved in the early period of vascular remodeling. The knowledge gained will suggest strategies for developing the best therapies for treatment of different vascular disorders.