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目的 探讨流场下E-选择素介导的中性粒细胞钙响应过程.方法 采用平行平板流动腔结合荧光显微镜,在0～600mPa流体剪应力(fluid shear stress,FSS)下,实时观察中性粒细胞在不同浓度E-选择素上的黏附及随后产生的钙响应.结果 在流场环境下E-选择素可特异性介导中性粒细胞的稳定黏附和钙响应,细胞的黏附数和激活比率随浓度的提高而逐渐增加.只有固定的E-选择素才能传递力信号来有效触发细胞的钙响应,FSS的增加不仅提高了细胞的激活比率(从23％增至70％),增加了钙响应的强度(从0.92增至1.45),而且大大缩短了细胞自黏附到钙响应启动的时间(从70s减为27s).结论 FSS和E-选择素协同调控中性粒细胞的钙响应,FSS对钙响应的速度和水平实行正向调节.研究结果可深化血流环境下白细胞免疫响应过程的理解.“,”Objective To investigate the E-selectin mediated-calcium response of neutrophils under flow fields. Methods A parallel-plate flow chamber combined with a fluorescence microscope was used to observe the adhesion and subsequent calcium response of neutrophils on E-selectin at different concentrations under fluid shear stress (FSS) of 0-600 mPa. Results E-selectin could capture neutrophils from the flow to the chamber substrate and induce further intracellular calcium flux of firmly adhered cells. The arrested cell and activation ratio increased gradually as the concentration of E-selectin increased. Only immobilized E-selectin could conduct the external force-signal to trigger the calcium response of neutrophils effectively. By increasing FSS, not only was the activation ratio increased from 23％ to 70％, but the calcium response intensity also increased from 0.92 to 1.45, while the delay time from cell adhesion to calcium response was greatly reduced from 70 s to 27 s. Conclusions FSS can modulate the calcium response of neutrophils in cooperation with E-selectin, and it positively regulates the activation rate and level of calcium response. This study may deepen the understanding of the immune response of leukocytes under a hemodynamic environment.