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.

我们将受控水平的流体剪切应力应用于特殊设计的流动室中培养的内皮细胞（EC），并检查了细胞内Ca^<2+>浓度（[[Ca^<2+>]）和粘附分子表达的变化。在ecs的存在下，在ecs ecs的存在下受到剪切应力，表现出剪切应力，显示出<2+ <2+ <2+ <2+ <2+ <2+ <2 ca^ca^ca^ca^ca^ca^ca^ca [ca^ca^ca^ca^ca^ca^ca]在单个细胞中观察到了不同的模式，包括峰值和高原，振荡和瞬态。该模式的百分比分别为62％，32％和6％。不管模式如何，[Ca^<2+>]从细胞边缘的特定基因座开始增加，并以Ca^<2+>波传播。该基因座对应于EC中富含小窝蛋白的基因座，这表明剪切应力的信息通过小窝进入。剪切应力抑制了小鼠淋巴结电网EC中血管粘附分子1（VCAM-1）的细胞表面表达，并同时降低了mRNA水平。 mRNA水平的降低是由于VCAM-1基因转录的抑制作用，VCAM-1启动子中的双AP1共有元件对于剪切诱导的转录活性抑制至关重要。这是一个负剪切应力响应元件，该研究首先证明了这一点。

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。”

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: "Fluid shear differentialy modulates adhesion molecule expression in human vascular endothelial cells.In "Organ Microcirculation:Bridging between Basic and Clinical Sciences",M.Tshuchiya et al.eds.," Excerpta Medica,Tokyo, 8 (1996)

J.Ando：“流体剪切差异调节人血管内皮细胞中的粘附分子表达。在“器官微循环：基础科学和临床科学之间的桥梁”中，M.Tshuchiya 等编辑，”Excerpta Medica，东京，8 (1996)

W.Yang: "Contribution of sustained Ca^<2+> elevation for nitric oxide production in endothelial cells and subsequent modulation of Ca^<2+> transient in vascular smooth muscle cells in coculture." J.Biol. Chem.271. 5647-5655 (1996)

W.Yang：“持续 Ca^2 升高对内皮细胞中一氧化氮的产生以及随后共培养中血管平滑肌细胞中 Ca^2 瞬时调节的贡献。”

J.Ando: "Flow-dependent regulation of gene expression in vascular endothelial cells." Jpn.Heart J., 14 (1996)

J.Ando：“血管内皮细胞中基因表达的流量依赖性调节。”