红树林淡紫拟青霉胞外多糖对小鼠DCs表型成熟的影响

Effects of extracellular polysaccharides from Paecilomyces Lilacinuson on function of mouse bone marrow-derived dendritic cells

目的:探讨红树林来源的淡紫拟青霉胞外多糖对小鼠骨髓源性树突状细胞(DCs)表型成熟的影响。方法:从小鼠骨髓腔中分离获得单核细胞,加入细胞因子rmG M-CSF、rmI L-4诱导分化为未成熟DCs并用不同浓度淡紫拟青霉胞外多糖干预,流式细胞术检测成熟DCs的表面标志CD11c、MHCⅡ、CD80、CD86分子的表达及吞噬FITC-dextran的能力,ELISA检测该多糖对DCs分泌IL-12的影响;MTT法检测该多糖对DCs刺激T细胞增殖的影响。结果:经400μg/ml多糖作用48 h后DCs表面分子CD11c、MHCⅡ、CD80、CD86的表达显著上调,与空白对照组相比P<0.01,与LPS组相比P>0.05;经该多糖作用后DCs吞噬FITC-dextran的能力下降,尤其300μg/ml、400μg/ml的多糖作用效果最明显,与空白对照组相比P<0.05;该多糖还可刺激DCs分泌IL-12,尤其100~400μg/ml的多糖刺激作用较强,与空白对照组相比P<0.05;另外,经该多糖处理的DCs还可刺激T细胞增殖,将不同比例刺激细胞与T作用后均可见T细胞增殖,与空白对照组相比P<0.05。结论:淡紫拟青霉胞外多糖可上调DCs表面CD11c、MHCⅡ、CD80和CD86分子表达,降低其吞噬能力,促进其分泌IL-12,还可刺激T细胞增殖,初步表明该多糖可刺激DCs分化成熟。

Objective： To investigate the effects of the Paecilomyces Lilacinuson extracellular polysaccharides on the phenotypic and maturation of murine dendritic cells. Methods： Imature DCs were induced in vitro from the murine bone marrow cells in the presence of rmG M-CSF and rmI L-4,and then they were cultured with different dosage of the extracellular polysaccharides. The morphological characterization was analyzed under microscopy. The expressions of the DCs surface costimulating factors and phagocytic function to FITC-dextran were detected by flow cytometry. The level of IL-12 secreted by DCs was observed by ELISA. At the same time the influence of DCs on the proliferation of T cells was determined by MTT. Results： Treating with the polysaccharides for 48 h could up-regulate the expression of DCs surface molecules,such as CD11 c,MHC Ⅱ,CD80 and CD86,and the 400 μg / ml was the optimal dose,comparing with the blank control group,the difference was significant（ P〈0.01）,contrast to LPS control group that was not different（ P〈0. 05）. The uptaking FITC-dextran ability of the DCs treated with 300 μg / ml and 400 μg / ml polysaccharides was significant lower than the unstimulated DCs（ P〈0.05）. At the same time the extract at different concentration could distinctly enhance the proliferation of T cells by DCs too. Conclusion： The extracellular polysaccharides could stimulate the maturation of dendritic cells and induce the production of mature dendritic cells.