青钱柳多糖对小鼠骨髓来源树突状细胞表面分子表达的影响

Effect of Cyclocarya paliurus (Batal.) Polysaccharides on the Surface Molecule Expression Level of Dendritic Cells

为探讨青钱柳多糖(CPC)对小鼠骨髓来源树突状细胞(BMDCs)细胞形态及表面分子的影响,首先采用细胞因子诱导法,以贴壁法获得贴壁单核细胞,添加重组粒细胞-巨噬细胞集落刺激因子(rmGM-CSF)和重组白细胞介素-4(rmIL-4)进行体外诱导,倒置显微镜及透射电镜观察细胞形态的变化;流式细胞术检测培养第6 dDCs的表面标志CD80,CD86和MHC II类分子的表达的变化。经CPC刺激24 h后,采用流式细胞术检测DCs表面MHC II类分子表达的变化。结果发现:经rmGM-CSF和rmIL-4诱导获得的DCs随着培养时间的延长,细胞形态发生改变,逐渐变成具有树状突起的DCs。经LPS刺激后的DCs能够发生典型的DCs的成熟,而培养第6天的DCs具有典型的DCs表型特征,可用于后续进一步实验。与阴性对照相比,CPC显著促进树突状细胞表面MHC II的表达,在浓度(10～200μg/mL)范围内呈现剂量依赖性。初步表明CPC可以促进树突状细胞的成熟。

To observe the effect of Cyclocarya paliurus （Batal.）polysaccharides （CPC）on differentiation and development of murine bone marrow derived dendritic cells（DCs） in vitro. Monocytes generated from Balb/e mouse were cultured in completed RPMI1640 medium containing 10% FBS, rmGM-CSF and rm IL-4. The morphology development was observed by invented microscope and transmission electron microscope. DCs on the 6th day were analyzed by flow cytometry （FCM） to determine the expression levels of CD50 ,CD86 and MHC Ⅱ. CPC was added to cells on day 6 of culture for 24 h, then the surface molecules expression level of MHC Ⅱ on DCs were analysis by FCM. The results showed that DCs induced by rmGM-CSF and rm IL-4 possessed typical morphology and phenotype on the 6 day,which can be used in the following experiments. Compared to untreated DCs, DCs treated with CPC increased the expression leves of MHC Ⅱ, which indicated that CPC could promote the phenotypic maturation of DCs.