黄芪多糖诱导脐血单核细胞向树突状细胞分化中的作用  

The role of NF-κB in the differentiation process of dendritic cells induced by astragalm polysaccharides from human cord blood monocyte

作　　者：陈志明[1] 邓旻[1] 朱仕兵[1] 隋向前[1] 李晓玲[1] 张晓洁[1] 徐望阳[1] 窦晓兵[2] 史亦谦[2] 

机构地区：[1]浙江中医药大学附属中西医结合医院,杭州310003 [2]浙江中医药大学

出　　处：《中华微生物学和免疫学杂志》2011年第4期305-311,共7页Chinese Journal of Microbiology and Immunology

基　　金：浙江省中医药管理局科研基金项目（2008CB059）;杭州市卫生局科研基金资助项目（2007A017）

摘　　要：目的观察NF-κB在黄芪多糖（APS）诱导脐血单核细胞向树突状细胞（DCs）分化过程的作用，探讨APS诱生DCs过程中的信号传导通路。方法无菌条件下采集脐血，密度梯度离心法获得脐血单核细胞分为3组。对照组：在无药物的RPMI 1640完全培养液中培养；APS组：在含有黄芪多糖100mg／L的RPMI1640完全培养液中培养；PDTC组：用NF-κB的抑制剂吡咯烷二硫氨基甲酸酯（PDTC）10μmol／L处理脐血单核细胞30min后，加入含有黄芪多糖100mg／L的RPMI1640完全培养液中培养。培养过程中用倒置光学显微镜和透射电镜观察细胞形态变化；收集培养第12天的细胞，FCM检测细胞免疫表型；免疫荧光显微镜下观察细胞内NF-κB的激活、迁移情况。结果（1）对照组细胞无成簇生长，培养至第12天细胞呈梭形巨噬细胞形态；黄芪多糖组细胞成簇生长，形态学由圆形逐渐变为典型的树突状细胞形态；抑制剂组细胞生长缓慢，未出现聚集现象，细胞形态学未见明显变化。（2）APS组高表达DCs特异性抗原CDSO、CD83和CD86，与对照组、PDTC组比较差异有统计学意义（P〈0．01），对照组与PDTC组表达相似，二者差异无统计学意义（P〉0．05）。（3）免疫荧光显微镜下观察NF-κB可见APs组伴有大量NF-κB荧光进入细胞核，尤以72h显著，NF-κB激活率为（75．20±7．37）％，而对照组、PDTC组NF-κB激活率分别为（13．20±3．46）％、（8．20±1．92）％，与APS组相比，差异有统计学意义（P〈0．02）。结论黄芪多糖能够诱导脐血单核细胞定向分化为DCs，NF-κB是黄芪多糖诱导脐血单核细胞向DCs分化的信号传导通路中的关键元件。Objective To investigate the role of NF-κB played in the process of the cord blood monoeytcs differentiating into dendritic cells（DCs） induced by astragalus polysacchafide（APS） and to explore the signal transduction pathway involved in this process. Methods Umbilical cord blood was collected in aseptic conditions. The cord blood monocytes were obtained by density gradient centrifugation and were divided into three groups afterwards. In the control group, cells were cultivated in the RPMI 1640 complete medium. In the APS group, cells were cultivated in the RPMI 1640 complete medium containing 100 mg/L APS. In the PDTC group： cells were treated with 10μmol/L disulfide carbamate（ PDTC）, NF-κB inlfibitor in 30 rain followed by cultivation in the RPMI 1640 complete medium containing 100 mg/L APS. The morphological changes were ob- served during the process of cultivation by the optical microscope and transmisaion electron microscopy. Cells were collected 12 d later and the cellular immunophenotyping was assayed by FCM. The activation and migration of NF-κB fluorescence in the cells was examined by the immunollourescence microscopy. Results （ 1 ） Cells in the control group grown up without cluster formation and were found fusiform and macroplmge-like in 12 d. Cells in the APS group grown up in clusters, and morphological changes were found from the circular shape to a typical dendritic cells-like shape. Cells in the inhibitor group grown up slowly and without cluster formation, and cell morphology had no significant change. （2） The expression of DCs--specific antigen CDSO, CD83 and CD86 in the APS group was higher than that in the control group and inhibitors group（ P 〈 0. 01 ）. The expression of those antigen in the control group and PDTC group was similar and had no statistically significance（ P 〉 0.05 ）. （3） NF-κB fluorescence in the nuclei was examined by the immunoflourescence microscopy and was much higher in the APS group than that in the other groups, especially in 72

关 键 词：脐血 树突状细胞 黄芪多糖/药理学 信号传导 

分 类 号：R285[医药卫生—中药学]