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目的比较血管内皮细胞(endothelial cells,ECs)在不同流场中转录因子核因子-кB(nuclear factor-кB,NF-кB)和激活蛋白-1(activator protein-1,AP-1)的表达规律,为力学信号对基因调控模式的影响提供依据。方法将培养的人脐静脉ECs加载于层流和湍流的离体流室模型中,以作用时程的不同分为6个时间点(0.5 h、1 h、2 h、3 h、4 h和5 h),运用共聚焦显微镜,从蛋白水平观察ECs中NF-кB和AP-1在不同流室中表达的时间规律和空间规律。结果层流中NF-кB的表达高峰出现在1 h段(26.49±1.63,P<0.05),随后转行向下;湍流中NF-кB表达在3 h段升至峰值(34.41±6.43,P<0.05);层流中c-Jun/AP-1表达一过性增高,0.5 h到达峰值(18.95±5.38,P<0.05),随即下降至基线水平;湍流中c-Jun/AP-1表达呈持续缓慢上升趋势(P<0.05)。结论湍流流场对血管ECs中NF-кB和AP-1表达的影响不同于层流,湍流对NF-кB和AP-1的持续激活上调作用可能引起了ECs形态结构和功能行为的病理学改变。
Objective To compare the expression of nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1) in vascular endothelial cells (ECs) , Provide the basis for the influence of mechanic signal on gene regulation mode. Methods Human umbilical vein endothelial cells (ECs) were cultured in laminar flow and turbulent flow chamber models. The time course of action was divided into 6 time points (0.5 h, 1 h, 2 h, 3 h, 4 h and 5 h). Confocal microscopy was used to observe the temporal and spatial rules of NF-кB and AP-1 expression in different flow compartments from the protein level. Results The peak expression of NF-кB in laminar flow occurred at 1 h (26.49 ± 1.63, P <0.05), and then turned downward. The expression of NF-кB in turbulence peaked at 3 h (34.41 ± 6.43, P < 0.05). The expression of c-Jun / AP-1 in laminar flow increased transiently and peaked at 0.5 h (18.95 ± 5.38, P <0.05), then decreased to the baseline level. Sustained slow upward trend (P <0.05). Conclusions The effect of turbulent flow on the expression of NF-кB and AP-1 in vascular ECs is different from that of laminar flow. The up-regulation of NF-кB and AP-1 by turbulence may cause the pathological changes of ECs morphology and function .