Mechanical regulation of calcium signaling of HL-60 on P-selectin under flow.
Mechanical regulation of calcium signaling of HL-60 on P-selectin under flow.
复制标题
流动下 HL-60 对 P-选择素的钙信号传导的机械调节。
DOI：
10.1186/s12938-016-0271-1
复制
发表时间：
2016-12-28
影响因子：
3.9
通讯作者：
Wu J
中科院分区：
文献类型：
--
作者：
Huang B;Ling Y;Lin J;Fang Y;Wu J
研究背景P-选择素与P-选择素糖蛋白配体-1（PSGL-1）的结合使中性粒细胞粘附于炎症部位。粘附的中性粒细胞的细胞内钙爆发是随后在损伤组织处牢固地捕获和迁移到损伤组织中的关键事件。但是，目前尚不清楚流体剪切力如何调节细胞的胞浆钙信号。在这里，我们专注于机械调节P-选择素诱导的Ca 2+信号的HL-60细胞在flow.MethodsHL-60细胞负载Fluo-4 AM荧光检测细胞内钙离子，然后灌注在P-选择素包被的平行板流动室的底部。荧光显微镜下真实的观察贴壁细胞内钙离子浓度的变化。结果力触发、增强和加速HL-60细胞对P-选择素的胞浆钙爆发。这种力依赖性钙信号传导是由在不存在趋化因子的情况下包被在基质上的固定化P-选择素诱导的。结论固定化P-选择素诱导的HL-60细胞钙信号转导具有P-选择素浓度依赖性和机械力依赖性。P-选择素浓度越高，细胞受到的外力越大，钙信号越强。这可能为粘附分子诱导的细胞内信号通路的机械化学调控机制提供新的见解。
BackgroundBinding of P-selectin to P-selectin glycoprotein ligand-1 (PSGL-1) makes neutrophils roll on and adhere to inflammatory site. Intracellular calcium bursting of adhered neutrophils is a key event for subsequent arresting firmly at and migrating into the injured tissue. But, it remains unclear how the cytoplasmic calcium signaling of the cells were modulated by the fluid shear stress. Here, we focus on mechanical regulation of P-selectin-induced calcium signaling of neutrophil-like HL-60 cells under flow.MethodsHL-60 cells were loaded with Fluo-4 AM for fluorescent detection of intracellular calcium ion, and then perfused over P-selectin-coated bottom of parallel-plate flow chamber. The intracellular calcium concentration of firmly adhered cell under flow was observed in real time by fluorescence microscopy.ResultsForce triggered, enhanced and quickened cytoplasmic calcium bursting of HL-60 on P-selectin. This force-dependent calcium signaling was induced by the immobilized P-selectin coated on substrates in absence of chemokine. Increasing of both shear stress and P-selectin concentration made the calcium signaling intensive, through quickening the cytosolic calcium release and upregulating both probability and peak level of calcium signaling.ConclusionsImmobilized P-selectin-induced calcium signaling of HL-60 cells is P-selectin concentration- and mechanical force-dependent. The higher both the P-selectin concentration and the external force on cell, the more intensive the calcium signaling. It might provide a novel insight into the mechano-chemical regulation mechanism for intracellular signaling pathways induced by adhesion molecules.