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目的研究不同血流状态下培养内皮细胞紧密连接的变化。方法设计一种新的平行平板流室模型,高速摄像微粒子示踪技术流体分析,培养内皮细胞进行7H6免疫荧光染色,F-肌动蛋白荧光染色和ZO-1双重免疫荧光染色,激光共聚焦显微镜观察。结果液体在入口与狭窄开始处中点的流速为249.8mm/sec,剪切应力为71.9dyn/cm2;进入狭窄段时的流速为316mm/sec,剪切应力为88.5dyn/cm2,上述区域液体为定常流。F-肌动蛋白纤维表达明显增加,呈束状,与液流方向一致,细胞变为细长。ZO-1和7H6则位于细胞周围呈现连续、线状环形分布。液体进入宽广区域后液体流速明显减慢并在两侧形成涡流甚至停滞,流速为54.4～119.6mm/sec,剪切应力为15.6～34.4dyn/cm2,F-肌动蛋白纤维分布和排列无方向性,细胞轮廓成不规则多边形,ZO-1和7H6在细胞周围呈点状分布。结论低剪切应力更易导致动脉硬化。
Objective To study the changes of endothelial cell tight junctions under different blood flow states. Methods A new parallel plate flow chamber model was designed. High-speed microphotography tracer technique was used to analyze the fluid and endothelial cells were cultured for 7H6 immunofluorescence staining, F-actin fluorescence staining and ZO-1 double immunofluorescence staining. Confocal laser scanning microscopy Observed. As a result, the flow rate of the liquid at the midpoint between the entrance and the stenosis was 249.8 mm / sec and the shear stress was 71.9 dyn / cm2; the flow rate at entry into the stenosis was 316 mm / sec and the shear stress was 88.5 dyn / cm2, For steady flow. F-actin fibers increased significantly, showing a bundle, consistent with the flow direction, the cells become slender. ZO-1 and 7H6 are located in the cell around the continuous, linear ring distribution. After the liquid enters the wide area, the flow velocity of the liquid slows down significantly and vortexes even stagnate on both sides. The flow velocity is 54.4-119.6 mm / sec and the shear stress is 15.6-34.4 dyn / cm2. The F-actin fibers are distributed and arranged without direction Sex, cell outline into irregular polygons, ZO-1 and 7H6 in the cell was dotted distribution. Conclusion Low shear stress is more likely to lead to atherosclerosis.