豚鼠腹腔巨噬细胞吞噬钩端螺旋体的机制研究  

Mechanisms of phagocytosis of Leptospira by peritoneal macrophages of guinea pigs

作　　者：娄晓丽[1] 张彦[1] 何平[1] 邓聪[2] 姜叙诚[2] 郭晓奎[1] 

机构地区：[1]上海交通大学基础医学院病原生物学教研室,上海200025 [2]上海交通大学基础医学院病理学教研室,上海200025

出　　处：《上海交通大学学报（医学版）》2009年第10期1143-1147,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家高技术研究发展计划("863"计划)(2006AA02Z176);国家自然科学基金(30770820);上海市自然科学基金(06ZR14056)~~

摘　　要：目的研究豚鼠腹腔巨噬细胞吞噬钩端螺旋体毒力株的机制,探讨天然免疫在钩端螺旋体病发病机制中的作用。方法提取豚鼠腹腔巨噬细胞,感染1 h前分别加入细胞微丝阻断剂cytochalasin D、微管阻断剂colchicine和PI3K信号通路阻断剂LY294002,同时设立不含阻断剂的对照组,1 h后检测细胞活性。与致病性问号钩体赖型Lai株共同孵育3 h后,激光共聚焦显微镜观察巨噬细胞吞噬钩体的形态学特征,流式细胞术检测各组吞噬率。结果对照组巨噬细胞对钩体的吞噬率为(38.98±0.91)%;cytochalasin D组、colchicine组和LY294002组的吞噬率分别为(23.99±1.40)%、(40.81±0.91)%和(39.64±3.56)%;cytochalasin D组吞噬率明显低于对照组(P<0.05);colchicine组和LY294002组与对照组比较差异无统计学意义(均P>0.05)。结论微丝参与了豚鼠腹腔巨噬细胞吞噬毒力钩体的过程,微管在吞噬过程中不起关键作用,微丝的变化不依赖PI3K信号通路。Objective To investigate the mechanisms of phagocytosis of virulent Leptospira by peritoneal macrophages of guinea pigs,and evaluate the role of innate immune in the pathogenesis of leptospirosis. Methods Peritoneal macrophages of guinea pigs were extracted.Three specific inhibitors（microfilament inhibitor cytochalasin D,microtube inhibitor colchicine and PI3K signalling pathway inhibitor LY294002） were added respectively to the macrophages 1 h before the infection of virulent Leptospira interrogans serovar Lai type strain Lai in vitro.Meanwhile,control group without inhibitor was established.Phagocytosis was observed by laser scanning confocal microscopy and phagocytic rates were evaluated by flow cytometry 3 h after infection. Results The phagocytic rates of control group,cytochalasin D group,colchicine group and LY294002 group were（38.98±0.91）%,（23.99±1.40）%,（40.81±0.91）% and（39.64±3.56）%,respectively.The phagocytic rate of cytochalasin D group was significantly lower than that of control group（P〈0.05）,while those of colchicine group and LY294002 group were not significantly different from that of control group（P〉0.05）.Conclusion Microfilaments play an important role in the phagocytosis of strain Lai by peritoneal macrophages,but the process is independent on PI3K signalling pathway,and microtubes play little part during the phagocytosis.

关 键 词：微管 微丝 PI3K 腹腔巨噬细胞 钩端螺旋体 

分 类 号：R514[医药卫生—内科学]