林珏龙1，陈耀文1，谢仰民2，李 玫2，朴仲贤1，杨海伟1
(1.汕头大学医学院中心实验室,中国广东 汕头 515031; 2.汕头大学医学院动物中心,
中国广东 汕头 515031)
摘 要：为评估小鼠巨噬细胞吞噬死亡细胞时胞质内游离钙离子的变化。实验使用Fluo-3标记巨噬细胞内钙离子和碘化丙碇对死亡细胞核染色，观察吞噬过程中细胞内钙离子的变化和显示巨噬细胞的吞噬功能，检测含死亡细胞的巨噬细胞内荧光密度图像。利用激光扫描共聚焦显微镜检测钙离子的释放。在缺钙的溶液中，可见巨噬细胞接触死细胞时细胞内钙离子快速地聚集和增高。在吞噬体形成时，巨噬细胞内钙离子上升到较高的水平。快速上升后，当吞噬小泡消化时，细胞内游离钙下降，随后钙离子恢复到低水平。研究显示伴随着吞噬小泡中红色荧光的死细胞的出现和消失，巨噬细胞内出现一系列钙离子变化的图像。提示巨噬细胞内钙离子改变在细胞吞噬作用中具有一定的作用。
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Cytosolic Free Clcium Fluctuation Mediates Phagocytosis in Mouse Macrophages
LIN Jue-long 1 , CHEN Yao-wen1 , XIE Yang-min 2 , LI Mei2 , PIAO Zhong-xian1 , YAN Hai-wei1
(1. Central Laboratory, Medical College, Shantou University, Shantou 515031, Guangdong,China;
2 Animal Centre, Medical College, Shantou University, Shantou 515041, Guangdong ,China )
Abstract : Cytosolic free calcium ([Ca2+]i ) change was assessed in single adherent mouse macrophages during phagocytosis for dead cells. To determine visualizing Ca2+ change during phagocytosis and indirectly show phagocytic function of macrophages to dead cells, intracellular [Ca2+]i of macrophages were loaded with the fluorescent dye Fluo-3/AM and dead cells were labeled with Propidium Iodide(PI), we examined images pattern of the fluorescence intensity in macrophages of containing dead cells. A discharge of calcium was verified by confocal scanning laser microscopy (CSLM). In lacking Ca2+ medium, upon contact with dead cells, rapid assemble and increase of [Ca2+]i was observed in macrophage. Intracellular [Ca2+]i rose to a higher level during initial phagosome formation. After a rapid rise, free Ca2+was decreased in digestion of phagocytotic vesicle; there-after the [Ca2+]i returned to the baseline. Our study directly indicates that accompany presenting and disappearing of dead cell of red fluorescence in the phagosomal vesicle, a series of images pattern of [Ca2+]i change can be really exhibited in macrophages. It is suggest that [Ca2+]i fluctuation has a certainty in macrophages during the phagocytosis.
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