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目的研究不同流体切应力对内皮祖细胞分泌组织纤溶酶原激活物(tissuetypeplasminogenactivator,tPA)的影响,探讨切应力调节内皮功能的作用。方法分离健康成人外周血的单个核细胞并诱导分化成内皮祖细胞,通过acLDL及lectin荧光抗体标记鉴定。种植一定数量内皮祖细胞到小径人工血管表面后,分为静态组、低切应力组(5dyn/cm2)、中切应力组(15dyn/cm2)和高切应力组(25dyn/cm2)4种不同处理组,用酶联免疫吸附法测定不同时间点(0、5、10、15、20、25h)培养液中tPA的水平。结果外周血单个核细胞分化成为内皮祖细胞,倒置荧光显微镜下呈典型的“纺锤样”梭形细胞,acLDL及lectin双抗体荧光标记阳性。流体切应力增加内皮祖细胞tPA的产生,tPA的合成与切应力的水平和时间有关。结论切应力调节内皮祖细胞tPA的分泌,提示流体切应力不仅是维持正常血管内皮功能的重要因素之一,也是动脉粥样硬化发病机制的重要环节之一。
Objective To investigate the effect of different fluid shear stress on the secretion of tissue plasminogen activator (tPA) by endothelial progenitor cells and to explore the role of shear stress in regulating endothelial function. Methods Mononuclear cells from healthy adult peripheral blood were isolated and induced to differentiate into endothelial progenitor cells, which were identified by acLDL and lectin fluorescent antibody labeling. After a certain number of EPCs were planted on the surface of small-diameter artificial blood vessels, they were divided into 4 groups: static group, low shear stress group (5dyn / cm2), middle shear stress group (15dyn / cm2) and high shear stress group (25dyn / cm2) In the treatment group, the level of tPA in the culture fluid at different time points (0, 5, 10, 15, 20, 25 h) was determined by enzyme-linked immunosorbent assay. Results Peripheral blood mononuclear cells differentiated into endothelial progenitor cells, which were typical “spindle” spindle cells under inverted fluorescence microscope. Fluorescent labeling of acLDL and lectin diabody was positive. Fluid shear stress increases the production of tPA in endothelial progenitor cells, and the synthesis of tPA correlates with the level and timing of shear stress. Conclusion Shear stress regulates the secretion of tPA in endothelial progenitor cells, suggesting that fluid shear stress is not only one of the important factors in the maintenance of normal vascular endothelial function, but also an important part of the pathogenesis of atherosclerosis.