钙激活钾通道KCa3.1对巨噬细胞吞噬功能的抑制性调节作用  

Inhibitory Effect of Ca^(2+)-activated Potassium Channel KCa31 on the Phagocytotic Function of RAW2647 Macrophages

作　　者：祝红[1,2] 刘枭[1,2] 李慧[1,2] 闫莉[1,2] 郝维[1,2] 曹济民[1,2] 

机构地区：[1]中国医学科学院基础医学研究所 [2]北京协和医学院基础学院生理学和病理生理学系,100005

出　　处：《医学研究杂志》2013年第7期21-24,76,共5页Journal of Medical Research

基　　金：国家科技部"973"基金资助项目(2011CB933500)"生物医学纳米材料对血细胞的作用研究"子课题4(2011CB933504)

摘　　要：目的观察巨噬细胞膜的钙激活钾通道KCa3.1与吞噬功能的关系。方法通过光镜下半定量观察RAW264.7巨噬细胞吞噬鸡红细胞,以及采用流式细胞术定量检测RAW264.7巨噬细胞吞噬异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的大肠杆菌(FITC-tagged E.coli)等方法,研究正常RAW264.7细胞的吞噬活动,以及阻断KCa3.1通道后RAW264.7细胞吞噬能力的改变。结果光镜检测结果显示,未受吞噬刺激因子激活的对照RAW264.7细胞对鸡红细胞有较弱的吞噬能力,其吞噬率为1.67%±0.29%;用KCa3.1通道选择性阻断剂TRAM-34(20nmol/L)处理RAW264.7细胞后,吞噬率增高至5.08%±0.38%,与对照组相比差异有统计学意义(P<0.01),且单个RAW264.7细胞吞噬鸡红细胞的个数也明显增加。流式细胞术结果显示,阻断KCa3.1通道后,RAW264.7细胞吞噬FITC标记的大肠杆菌的数量明显增多,与对照组相比差异有统计学意义(P<0.01)。结论 KCa3.1通道的活动对RAW264.7巨噬细胞的吞噬活性有抑制性调节作用。Objective To investigate the relationship between Ca2＋ - activated potassium channel KCa3. I activity and the phagocytot- ic function of macrophages. Methods Microscopy and flow cytometry were used to semi - quantitatively and quantitatively estimate the phagocytotic activities of murine macrophage cell line RAW264.7 on chicken red blood cells and FITC - tagged E. coli k - 12, with or without TRAM -34 （a KCa3.1 selective blocker） pre -treatment, respectively, and thus to determine the role of KCa3.1 in the phagocyto- sis. Results Under a light microscope, un excited RAW264.7 cells （control） showed that a weak phagocytotic activity on chicken red blood cells, the phagocytototic rate （PR） was only 1. 67% ＋ 0. 29%. However, pre - treatment of the RAW264. 7 cells with TRAM-34 （20nmol/L） significantly increased the PR to 5.08% ＋0.38% （P 〈0.01 vs control）. In the flow cytometry assay, blocking KCa3.1 with TRAM - 34 also significantly increased the phagocytotic activities of RAW264.7 cells on FITC - tagged E. coli k - 12 compared with the control （P 〈 0.01 ）. Conclusion The activity of KCa3.1 exerts an inhibitory role on the phagocytotic function of RAW264.7 macrophages.

关 键 词：钙激活钾通道 巨噬细胞 吞噬 鸡红细胞 大肠杆菌 

分 类 号：R285.5[医药卫生—中药学]