补体受体3在小鼠巨噬细胞识别马尔尼菲青霉中的作用  

Roles of complement receptor 3 on murine macrophages in recognition of Penicillium marneffei

作　　者：胡永轩[1,2] 张军民[1] 鲁莎[1] 李希清[1] 梁宇恒[1] 鲁长明[1] 席丽艳[1] 

机构地区：[1]中山大学孙逸仙纪念医院,广州510120 [2]南方医科大学第三附属医院,510630广州

出　　处：《中华皮肤科杂志》2013年第8期538-542,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金一广东联合基金（U0932003）;南方医科大学科研基金（2012046）

摘　　要：目的探讨补体受体3（CR3）在小鼠巨噬细胞识别马尔尼菲青霉中的作用。方法以小鼠巨噬细胞系RAW264．7为靶细胞，分别与马尔尼菲青霉灭活分生孢子、灭活酵母细胞、活分生孢子、活酵母细胞在37oC，5％CO2培养箱中共同培养1h后，逆转录PCR检测CR3mRNA的表达，Western印迹检测CR3蛋白的表达，流式细胞仪检测吞噬率，酶联免疫吸附试验（ELISA）检测共培养上清中细胞因子水平。siRNA靶向下调巨噬细胞CR3的表达后与马尔尼菲青霉灭活分生孢子共培养，按照前述方法检测吞噬率及各细胞因子水平。采用SPSS16．0统计软件进行单因素方差分析（ANOVA）检测各组间指标差异。结果RAW264．7细胞与马尔尼菲青霉灭活分生孢子、灭活酵母细胞、活分生孢子、活酵母细胞共培养后，四组之间比较，CR3mRNA、蛋白表达差异均无统计学意义（P〉0．05），吞噬率分别为95．14％、89．56％、91．03％、90．78％，各组之间差异无统计学意义（P〉0．05）；共培养后，白介素（IL）2、干扰素（IFN）γ等Th1型，IL-4、IL-10等Th2型细胞因子均呈不同程度升高，但四个共培养组之间差异无统计学意义（P〉0．05）。siRNA靶向下调CR3表达后，RAW264．7细胞与马尔尼菲青霉共培养，其对马尔尼菲青霉的吞噬率下降为10．89％，与未干扰组比较，差异有统计学意义（P〈0．05）；同时，巨噬细胞分泌细胞因子的水平与未干扰组比较下降。结论CR3为巨噬细胞识别、介导吞噬马尔尼菲青霉的模式识别受体之一；IL-2、IFN-γ Th1型，IL-4、IL-10Th2型细胞因子可能均参与巨噬细胞抗马尔尼菲青霉感染免疫。Objective To evaluate the role of complement receptor 3 （CR3） on murine macrophages in the recognition of Pcnicillium marneffei. Methods RAW264.7 murine macrophage cells were cultured in vitro, and divided into four groups to be cocuhured with inactivated and live Penieillium mameffei yeast cells as well as inactivated and live Penicillium marneffei conidia respectively at 37 ℃ in 5% CO2 for one hour. The RAW264.7 cells incubated with phosphate-buffered saline （PBS） served as the blank control group. Then, reverse transcription-PCR was conducted to detect CR3 mRNA expression, Western blot to measure CR3 protein expression, flow cytometry to determine phagocytosis rate, enzyme-linked immunosorbent assay （ELISA） to quantify cytokine levels in culture supernatant. Some RAW264.7 macrophages were transfected with a specific siRNA targeting CR3 gene and cocultured with inactivated Penicillium marneffei conidia, subsequently, phagocytosis rate and supernatant cytokine levels were determined. Data were processed by the SPSS 16.0 software, and one-way analysis of variance （ANOVA） was conducted for inter-group comparisons of these parameters. Results No significant differences were observed in the mRNA or protein expressions of CR3 among the four groups of RAW264.7 cells cocuhured with different forms of Penicillium marne/irei （both P 〉 0.05）. The phagocytosis rate was 95.14%, 89.56%, 91.03% and 90.78% in RAW264.7 cells cocuhured with inactivated conidia and yeast cells, as well as live conidia and yeast cells of Penicillium marneffei, respectively （P 〉 0.05）. The levels of interleukin （IL）-2, interferon （IFN）-γ IL-4 and IL-10 in culture supernatant were increased at different degrees after one-hour cocuhure in the four coculture groups compared with the blank control group, but no statistical difference was noted among the four cocuhure groups in the supernatant levels of these cytokines （all P 〉 0.05）. After cocuhure with inactivated Penicillium marneffei conidia, the siRNA-tr

关 键 词：巨噬细胞 马尔尼菲青霉 受体 补体 疾病模型 动物 

分 类 号：R379[医药卫生—病原生物学]