A Bio-Engineering Approach to Endothelial Cell Signal Transduction and Responses to Fluid Shear Stress

内皮细胞信号转导和流体剪切应力响应的生物工程方法

• 批准号: 07408036
• 负责人: KAMIYA Akira
• 金额: $ 1.79万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07408036/
• 关键词: SHEAR STRESS; ENDOTHELIAL CELL; CALCIUM ION; VCAM-1; ADHESION MOLECULE; BLOOD FLOW; HEMODYNAMIC FORCE; VCAM-1; 血管内皮細胞; 流れずり応力; 細胞内カルシウム; 血行力学的応力; 遺伝子; 細胞内情報伝達系

We applied controlled levels of fluid shear stress to cultured endothelial cells (ECs) in a specially designed flow-loading chamber, and examined the changes in intracellular Ca^<2+> concentrations ([Ca^<2+>]) and adhesion molecule expressions.When subjected to shear stress in the presence of extracellular ATP,ECs showed the increase in [Ca^<2+>]. Different patterns including peak & plateau, oscillation, and transient were observed in individual cells. The percentage of the pattern was 62%, 32%and6%, respectively. Regardless of the patterns, [Ca^<2+>] increase initiated from specific loci at cell edges, and propagated through the entire cell body as a Ca^<2+> wave. The loci corresponded to the caveolin-rich in ECs, suggesting that the information of shear stress enters via caveolae.Shear stress inhibited the cell surface expression of vascular adhesion molecule-1 (VCAM-1) in mouse lymphnode venule ECs, and simultaneously decreased the mRNA levels. The decrease in mRNA levels was due to the suppression of VCAM-1 gene transcription, and double AP1 consensus elements in VCAM-1 promoter was essential for the shear-induced suppression of transcriptional activity. This is a negative shear stress responsive element, which was first demonstrated by this study.

我们在一个专门设计的流动加载室中对培养的内皮细胞施加一定水平的流体剪切力，检测细胞内Ca^<2 +>浓度（[Ca ^<2 +]）和粘附分子表达的变化。在单个细胞中观察到不同的模式，包括峰和平台，振荡和瞬态。三种模式的发生率分别为62%、32%和6%。不管是哪种模式，[Ca^2+]的增加都是从细胞边缘的特定位点开始的，并以Ca^2+波的形式传播到整个细胞体。切应力可抑制小鼠淋巴结微静脉内皮细胞表面血管黏附分子1（VCAM-1）的表达，同时降低其mRNA水平。mRNA水平的降低是由于VCAM-1基因转录受到抑制，而VCAM-1启动子中的双AP 1共有元件是剪切诱导转录活性抑制所必需的。这是一个负剪应力响应元件，这是第一次证明了这项研究。

项目成果

W.Yang: "Exogenous nitric oxide inhibits proliferation of cultured vascular endothelial cells." Biochem.Boiphys.Res.Commun.203. 1160-1167 (1994)

W.Yang：“外源性一氧化氮抑制培养的血管内皮细胞的增殖。”

W. Yang: "Exogenous nitric oxide inhibits proliferation of cultured vascular endothelial cells." Biochem. Biophys. Res. Commun.203. 1160-1167 (1994)

W. Yang：“外源性一氧化氮抑制培养的血管内皮细胞的增殖。”

Y.Takada: "Fluid shear stress increases the expression of thrombomodulin by cultured human endothelial cells." Biochem.Biophys.Res.Commun.205. 1345-1352 (1994)

Y.Takada：“流体剪切应力增加了培养的人内皮细胞的血栓调节蛋白表达。”

Y.Takada: "Fluid shear stress increases the expression of thrombomodulin by cultured human endothelial cells." Biochem.Boiphys.Res.Commun.205. 1345-1352 (1994)

Y.Takada：“流体剪切应力增加了培养的人内皮细胞的血栓调节蛋白表达。”

J. Ando: "Shear stress inhibits the adhesion of cultured mouse endothelial cells to lymphocytes by down-regulating VCAM-1 expression." Am. J. Physiol.267. C679-C687 (1994)

J. Ando：“剪切应力通过下调 VCAM-1 表达来抑制培养的小鼠内皮细胞与淋巴细胞的粘附。”