【摘要】 目的研究透明质酸(HA)对巨噬细胞吞噬功能的影响及其机制。方法用免疫细胞化学法标记CD44。在噬动轨迹实验中,用图像分析仪测量细胞吞噬后在其周围形成的无颗粒区面积,扫描电镜下观察巨噬细胞形态。在吞噬荧光微珠实验中,计数吞噬的微珠,观察细胞内F-肌动蛋白的分布。在吞噬HA包被微珠实验中,计数吞噬的微珠,在扫描电镜下观察细胞形态,用激光扫描共焦显微镜分析吞噬HA包被微珠时细胞内Ca2+水平。用CD44阻断抗体处理后,观察细胞吞噬功能的变化。结果巨噬细胞表达CD44。吞噬细胞较圆,板状伪足和丝状伪足较短。在吞噬颗粒和微珠时,细胞膜出现杯状凹陷。在HA组,吞噬氯化金颗粒和荧光微珠的数目增多,吞噬荧光微珠处的F-肌动蛋白丰富。在包被HA微珠组,吞噬微珠的数目增多,吞噬处的Ca2+水平显著升高。阻断CD44后,细胞吞噬氯化金颗粒、荧光微珠和HA包被微珠减少。结论HA促进巨噬细胞的吞噬功能,CD44介导HA对巨噬细胞吞噬功能的作用。更多还原

【Abstract】 Objective To investigate the effects of hyaluronan（HA） on phagocytosis of macrophages and to explore the mechanism.Methods CD44 of macrophages was labeled with immunochemical method.In phagokinetic track assay,the areas without gold particles around the cells were measured with an image analyzer,shape of the cells was viewed under a scanning electron microscope.In fluorescent bead phagocytosis assay,the phagocytosed beads were conunted,the distribution of intracellular F-actin was viewed.In HA-coated bead phagocytosis assay,the phagocytosed beads were counted,shape of the cells was viewed with scanning electron microscope,the level of the intracellular Ca²⁺ was analyzed with confocal laser scanning microscope when the beads were phagocytosed.After treatment with anti-CD44 antibody,the changes of phagocytosis of the cells were examined.Results CD44 expressed on macrophages.The phagocytosing cells were round,their lamellipodia and filopodia were short.The membrane of the cells formed cup-like invagination when they engulfed particles and beads.In HA group,the phagocytosed gold particles and fluorescent beads increased.There was rich F-actin in the area where the fluorescent bead was engulfed.In HA-coated group,number of the phagocytosed HA-coated beads was great,the level of the intracellular Ca²⁺ in the area where the bead was engulfed increased significantly.After treatment with anti-CD44 antibody,the phagocytosed gold particles,fluorescent beads and HA-coated beads decreased.Conclusion HA promotes phagocytosis of macrophages,CD44 mediates effects of HA on phagocytosis of the cells.更多还原