壳寡糖结合并激活巨噬细胞机制的研究  

Binding and stimulatory effect of oligochitosan in macrophages

作　　者：侯丽娜[1] 赵鲁杭[1] 

机构地区：[1]浙江大学医学院生物化学与分子生物学教研室,浙江杭州310031

出　　处：《中国医科大学学报》2006年第2期124-127,共4页Journal of China Medical University

基　　金：浙江省自然科学基金资助项目(302031)

摘　　要：目的:研究壳寡糖结合并激活巨噬细胞的机制。方法:RT-PCR和ELISA检测壳寡糖对巨噬细胞及经γ干扰素(IFN-γ)预刺激巨噬细胞的白介素1β(IL-1β)基因表达水平的影响;钙离子脂化探针F luo-3/AM检测壳寡糖对巨噬细胞胞浆游离Ca2+浓度的影响;用四甲基异硫氰酸罗丹明标记甘露聚糖,激光共聚焦显微镜和流式细胞仪观察标记糖结合及进入巨噬细胞的情况;通过竞争性抑制实验探讨巨噬细胞摄取壳寡糖的可能途径。结果:壳寡糖对巨噬细胞IL-1β基因表达的影响呈浓度与时间依赖性,IFN-γ预刺激巨噬细胞有增强作用;高浓度壳寡糖引起巨噬细胞胞浆游离Ca2+浓度升高;壳寡糖竞争性抑制巨噬细胞内吞甘露聚糖,抑制强度达44%。结论:壳寡糖可能是经由巨噬细胞表面的甘露糖受体介导结合并激活巨噬细胞,钙离子相关的信号转导途径可能参与这一过程。Objective： To explore the mechanism of the binding and stimulatory effect of oligochitosan in macrophages （MP）. Methods： The effect of oligochitosan on interleukin-1β（IL-1β） gene expression in resting macrophages and macrophages prestimulated with interferon-γ （IFN-γ ） was detected by using reverse transcription polymerase chain reaction （RT-PCR） and enzyme-linked immunosorbent assay （ELISA）. Macrophages treated with oligochitosan of different concentrations were dyed with Fluo-3/AM to observe the effect of oligochitosan on the level of free Ca2^＋ in the cytoplasm. Mannan was labeled with tetramethyl rhodamine isothiocyanate. The interactions between macrophages and the labeled sugar were observed under confocal laser microscope and analyzed by using flow cytometry. Competitive inhibition study was performed to determine the mechanism of oligochitosan uptake by macrophages. Results： Oligochitosan stimulated IL-1β gene expression in the concentration-dependent and time-dependent manner, and this stimulatory effect was augmented by the prestimulation with IFN-γ. Oligochitosan of high concentration increased the level of free Ca2^＋ in the cytoplasm. Oligochitosan competitively inhibited the uptake of mannan in macrophages, and the inhibitory intensity was as high as 44 %. Conclusion. Oligochitosan may bind and stimulate macrophages via mannose receptor, and some Ca2^＋ dependent signaling pathway may participate in this process.

关 键 词：壳寡糖 巨噬细胞 甘露糖受体 白细胞介素1p 1干扰素 

分 类 号：R378.911[医药卫生—病原生物学]