莫诺苷对小鼠骨髓源树突状细胞表型及功能的影响

Effects of morroniside on the properties of bone marrow derived dendritic cells

目的:探究莫诺苷对小鼠骨髓源树突状细胞(bone-marrow derived dendritic cells,BMDCs)分化、表型和功能的影响。方法:体外培养C57BL/6小鼠BMDCs,分为对照组(RPMI 1640)、莫诺苷组、脂多糖组和莫诺苷+脂多糖联合处理组。用光学显微镜以及流式细胞术分别确认BMDCs形态变化和羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)的荧光强度;流式细胞术检测BMDCs表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ类分子的表达情况以及激活抗原特异性CD4^+T细胞活化、增殖和分化的能力;ELISA法检测BMDCs培养上清液中细胞因子IL-6和IL-12的含量。结果:与对照组相比,莫诺苷处理后BMDCs诱导分化过程中的增殖效率无明显差异;与脂多糖组相比,莫诺苷+脂多糖联合处理后上调成熟BMDCs表面CD80、CD86、MHC-Ⅰ、MHC-Ⅱ类分子的表达和IL-12的分泌,并促进BMDCs激活OVA323-339抗原特异性T细胞的活化、增殖以及向Th1分化。结论:莫诺苷促进小鼠BMDCs的成熟,上调其对OVA特异性CD4^+T细胞的抗原提呈功能。

Objective: To investigate the effects of morroniside on the differentiation,phenotype and function of murine bone-marrow derived dendritic cells( BMDCs) in vitro. Methods: Primary murine BMDCs were prepared and divided into four groups: normal control group,morroniside-treated group,LPS-treated group and morroniside + LPS-treated group. The changes of BMDCs morphology and fluorescence intensity of CFSE were confirmed respectively with light microscopy and flow cytometry( FCM).The phenotype and functional maturation of BMDCs and activation of antigen-specific CD4^+T cell proliferation and differentiation under influence of morroniside were evaluated by FCM. ELISA was used for analyzing the protein levels of IL-6/IL-12 in supernatant of cultured BMDCs. Results: Compared with the normal control group,there was no significant difference in the proliferation efficiency of BMDCs induced differentiation after morroniside treatment. Compared with the LPS-treated group,morroniside combined with LPS could up-regulated the expression of CD80,CD86,MHC-Ⅰ and MHC-Ⅱ molecules and promoted the IL-12 secretion on BMDCs. Furthermore,morroniside treatment promoted activation,proliferation and differentiation priming capability on OVA323-339 antigen-specific T cells of BMDCs. Conclusion: Morroniside effectively enhances the maturation of BMDCs and up-regulates the antigen-presenting function of OVA-specific CD4^+T cells in vitro.