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ATP活化的小鼠巨噬细胞通讯功能的研究 |
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吕桂芝 张志谦 林仲翔
【摘要】 目的 探索间隙连接通讯功能、间隙连接蛋白CX43表达、微丝、纽蛋白在巨噬细胞活化过程中所起的重要作用。 方法 ATP作用于由小鼠腹腔获取的巨噬细胞(Mφ)，用MTT法检测ATP对巨噬细胞的活化作用，用罗氏黄(Ly)荧光染料示踪术检测ATP对小鼠腹腔巨噬细胞间隙连接通讯功能的影响，用免疫荧光技术检测ATP作用下的Mφ连接蛋白Cx43的表达和粘着斑-纽蛋白表达的影响，用罗丹明-鬼笔环肽-F-actin专一标记技术，观察ATP作用下的Mφ微丝蛋白(F-actin)的变化。 结果 (1)ATP对Mφ有明显的活化作用；(2)ATP上调Mφ的间隙连接通讯功能；(3)ATP增强连接蛋白Cx43的表达；(4)ATP增强粘着斑主要粘着蛋白-纽蛋白的表达；(5)ATP改善Mφ微丝束的组装。 结论 在ATP促进Mφ的活化过程中，间隙连接通讯功能、连接蛋白、纽蛋白、微丝蛋白都参与了Mφ活化过程中信号传导的调节作用。
【关键词】 ATP 巨噬细胞 活化作用 间隙连接通讯功能 连接蛋白 纽蛋白 微丝蛋白
INVESTIGATION OF COMMUNICATED FUNCTION ON ATP
ACTIVATED MACROPHAGES
Lü Guizhi△, Zhang Zhiqian, Lin Zhongxiang
(Beijing Institute for Cancer Research, School of Oncology, Beijing Medical University, Beijing)
【Abstract】 Objective In order to investigate the major effects of gap junctional intercellular communication(GJIC),connexin(CX43) and cytoskeleton protein(vinculin and F-actin) expression in ATP activated Mφ process. Methods Mouse peritoneal macrophages (Mφ)were used in this work. Experimental group (Mφ) was added with ATP into peritoneal Mφ solution, control group was used with peritoneal Mφ solution. Two group s Mφ cells were seeded in 96 well tissue culture plate or glass slide respectively and cultured in CO2 incubator 37℃ for 15 , activity of Mφ cells were examined by using MTT method. Expression of CX43 was demonstrated by using immunofluorescent cytochemistry method, GJIC was assessed by examining transfer of lucifer yellow dye after loading in the medium, microfilament cytoskeleton was studied by using double staining with rhodamine-phalloidin for F-actin and immunofluorescence for vinculin. Results It was found that (1) ATP stimulated Mφ cells obviously.(2) Increase of GJIC was observed in ATP-stimulated Mφ cells.(3) CX43 expression of ATP-stimulated Mφ cells was increased. (4) Improved of organization of vinculin positive adhesion plaqu[1] [2] [3] 下一页
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