EFFECTS OF FLOW & PRESSURE ON CULTURED ENDOTHELIAL CELLS

流动的影响

• 批准号: 3337119
• 负责人: SUZANNE G ESKIN
• 金额: $ 17.4万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-05-01 至 1991-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3337119
• 关键词: actins; blood pressure; cell adhesion; cell migration; cinemicrography; eicosanoid metabolism; electron microscopy; extracellular matrix; fluorescence spectrometry; gap junctions; hemodynamics; hemostasis; human tissue; immunofluorescence technique; inflammation; inositol phosphates; mechanical stress; membrane proteins; physical model; radioimmunoassay; scanning electron microscopy; thrombosis; tissue /cell culture; vascular endothelium; vascular endothelium permeability; vinculin

To develop effective preventative and therapeutic regimens for cardiovascular disease, it is important to understand control of he functional state of the vascular endothelial cells (EC). EC are naturally subjected to physical forces in situ. Thus definition of the precise roles of physical forces in maintaining normal and in producing pathological EC behavior is important. Shear stress, the force tangential to the EC resulting from blood flow is considered to be the mechanical force that is most injurious to the endothelium. Whether pulsatile variations in shear stress differ as a stimulus from steady shear stress is unknown. This research will analyze the mechanisms of adaptive EC responses to steady shear stress, and compare these with responses to pulsatile shear stress. The specific aims are to 1. Determine the morphologic changes in EC adhesions as they remodel under shear stress, 2. Determine the composition and distribution of ablumenal adhesion plaque components in shear stressed EC, 3. Determine the distribution of lumenal surface membrane proteins under shear stress, 4. Determine the effects of shear stress on directional migration using a "wound" model, 5. Determine whether inositol lipid turnover and arachidonic acid metabolism are linked to the shear stress induced structural response of EC, and 6. Compare the effects of shear stress on EC cultured on a permeable substrate to those of EC cultured on conventional substrates. The following techniques will be used to accomplish the specific aims; digital analysis of video enhanced interference reflection and immunofluorescent live cells under shear stress, microinjection of fluorescently labelled cytoskeletal proteins and fluorescence photobleach recovery of incorporated protein, transmission electron microscopy, radioimmunoassays, and spectrofluorometry. The long term results of our basic cell biological and engineering studies will elucidate the mechanisms whereby endothelium carries out its functions of modulating hemostasis and thrombosis and controlling vascular tone and permeability. EC abnormalities may contribute to atherosclerosis, thrombosis, hemostatic disorders, as well as to inflammation, immune reactivity and tumorigenesis. How shear stress alters EC function will elucidate these disease processes.

制定有效的预防和治疗方案， 心血管疾病，重要的是要了解控制他 血管内皮细胞（EC）的功能状态。 当然，EC 在现场受到物理力的作用。 因此，准确的角色定义 在维持正常和产生病理性EC的物理力量 行为很重要。 剪切应力，与EC相切的力 血液流动产生的机械力被认为是 对内皮损伤最大。 是否剪切的脉动变化 应力作为刺激不同于稳定剪切应力是未知的。 这 研究将分析适应性EC响应稳定的机制， 剪切应力，并将其与脉动剪切应力响应进行比较。 具体目标是1. 确定EC的形态学变化 粘连，因为他们在剪切应力下重塑，2。 确定组成 切应力作用下牙槽骨粘附菌斑成分的分布 EC，3. 确定管腔表面膜蛋白的分布 在剪切应力下，4. 确定剪切应力对 使用“伤口”模型的定向迁移，5. 确定肌醇是否 脂质周转和花生四烯酸代谢与剪切力有关 应力诱导EC结构响应; 6. 疗效比较 在可渗透基质上培养EC上的剪切应力与EC的剪切应力相比 在常规基质上培养。 将使用以下技术 以实现具体目标;数字分析视频增强 剪切作用下肝细胞干涉反射与免疫荧光 应激，荧光标记的细胞骨架蛋白的显微注射， 掺入蛋白质的荧光光漂白恢复，透射 电子显微镜、放射免疫测定法和荧光分光光度法。 我们基础细胞生物学和工程学研究的长期结果 将阐明内皮细胞发挥其功能的机制 调节止血和血栓形成以及控制血管张力， 磁导率 EC异常可能导致动脉粥样硬化、血栓形成、止血 疾病，以及炎症、免疫反应性和肿瘤发生。 切应力如何改变EC功能将阐明这些疾病的过程。