Mechanisms of morphological and functional changes of endothelial cells in atherogesesis.

动脉粥样硬化中内皮细胞形态和功能变化的机制。

基本信息

批准号：
14570204
负责人：
MITSUMATA Masako
金额：
$ 2.37万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570204/
关键词：
atherosclerosis endothelial cells shear stress apoptosis cell proliferation monocyte adhesion MMP-2 en face method 増殖血流 p21 血管内皮面画像 IAP 血管内皮面立体画像

项目摘要

1.To disclose the anti-atherosclerotic mechanisms of steady laminar shear stress, we analyzed the expression of human inhibitor of apoptosis protein-2(HIAP-2) in endothelial cells(ECs). HIAP-2 was dose-dependently and time-dependently induced in ECs by shear stress. HIAP-2 expression also identified in vivo. Shear stress-mediated inhibition of EC apoptosis was associated with inhibition of caspase-3 activity. Transfection of Smac, a caspase activator by binding HIAPs and removing their inhibitory activity, reduced the suppression effect of shear stress on caspase-3 activity. These data suggest that shear stress prevents EC apoptosis via negative regulation of caspase-3 by increments of HIAP-2. 2.The MMP2 secretion from ECs into medium was interrupted completely by laminar shear stress, although level of this intracellular protein and mRNA expression of MMP2 were unchanged. To clarify this mechanism, we analyzed intracellular transportation of MMP-2, using fused MMP-2 tagged with GFP. B … More y laminar shear stress, MMP-2/GFP granules increased their size and transported into cell membrane without secretion. Our data suggest that laminar shear stress contribute to EC adhesion on matrix through the regulation of collagen type IV metabolism. 3.To analyze a initial events of atherosclerosis, in vivo, we established a modified en face method that enabled us to obtain clear images of the entire surface of endothelium including a bifurcation orifices of aorta, where atherosclerosis initiates. A number of proliferating ECs and macrophages adhered on ECs, stained with specific antibodies, at the branching area of aorta from SD rats were about 2 and 3 times higher than that of non-branching area, respectively. In conclusion, stabilization of ECs through the regulation of cell death, cell growth, EC adhesion on matrix with control of collagen metabolism and inhibition of monocyte adhesion on ECs by the laminar shear stress may contribute to the formation of the anti-atherosclerotic microenvironment in vessels. Less

1.揭示稳定层状剪切应力的抗动脉粥样硬化机制，我们分析了内皮细胞（ECS）中凋亡蛋白-2（HIAP-2）抑制剂的表达。剪切应力在EC中诱导HIAP-2剂量依赖性和时间依赖性。 HIAP-2表达也在体内鉴定。剪切应力介导的EC凋亡抑制与caspase-3活性的抑制有关。通过结合HIAP并去除其抑制活性，SMAC（SMAC）转染降低了剪切应力对caspase-3活性的抑制作用。这些数据表明，剪切应力通过caspase-3的负调控通过HIAP-2的增量来阻止EC凋亡。 2.尽管这种细胞内蛋白的水平和mMP2的mRNA表达不变，但从ECS分泌到培养基的MMP2分泌完全被层状剪切应力中断。为了阐明这种机制，我们使用用GFP标记的FISUDE MMP-2分析了MMP-2的细胞内转运。 B…更多的层层剪切应力，MMP-2/GFP颗粒增加了大小，并不分解进入细胞膜。我们的数据表明，层状剪切应力通过调节IV型代谢而导致基质上的EC粘附。 3.分析动脉粥样硬化的初始事件，在体内，我们建立了一种改进的面部方法，使我们能够获得整个山地座表面的清晰图像，包括主动脉粥样硬化的分叉顺序，动脉粥样硬化会引起动脉粥样硬化。来自SD大鼠主动脉的分支区域的EC上粘附的许多增殖EC和巨噬细胞分别比非分支区域的分别高约2和3倍。总之，通过调节细胞死亡，细胞生长，基质对胶原蛋白代谢的EC粘附的稳定，并通过层状剪切应力控制了胶原蛋白代谢并抑制单核细胞粘附于ECS上的ECS粘附，可能有助于形成船舶中抗抗溶质性微微环境。