神经生长因子在诱导树突状细胞分化中的作用

发布时间：2018-07-06 17:17

本文选题：神经生长因子 + 树突状细胞　；参考：《中南大学学报(医学版)》2015年08期

【摘要】：目的:在体外探讨神经生长因子(nerve growth factor,NGF)在刺激小鼠骨髓来源的树突状细胞(dendritic cells,DCs)分化过程中的作用。方法:首先取C57BL/6小鼠股骨、胫骨骨髓细胞,按培养过程中干预因素不同分为磷酸盐缓冲液(phosphate buff ered saline,PBS)刺激组(PBS组,n=6)、NGF刺激组(NGF组,n=6)、粒巨噬细胞集落刺激因子(granulocyte monocyte colony stimulating factor,GM-CSF)和IL-4共刺激组(GM-CSF+IL-4组,n=6)、NGF加GM-CSF和IL-4共刺激组(NGF+GM-CSF+IL-4组,n=6),培养第7天采用流式细胞仪比较各组间CD11c+细胞阳性率和CD8a-亚型比例;然后用GM-CSF+IL-4刺激培养未成熟的DCs,经CD11c免疫磁珠阳性分选得到纯化的未成熟DCs,将其分为PBS组(n=6)、NGF组(n=6)、脂多糖(lipopolysaccharide,LPS)组(LPS组,n=6)、NGF+LPS组(n=6),分别用PBS,NGF,LPS,NGF+LPS诱导细胞成熟,24 h后酶联免疫吸附分析法(enzyme linked immunosorbent assay,ELISA)检测上清液中IL-6,IL-10和IL-12的水平。结果:1)NGF组CD11c+DCs百分比高于PBS组,低于GM-CSF+IL-4组(均P0.05);GM-CSF+IL-4组与NGF+GM-CS F+IL-4组间比较,差异无统计学意义(P0.05),且在4组中均以CD8a-DCs为主;2)NGF可以增强LPS诱导的DCs分泌IL-6,IL-10和IL-12(均P0.05),而单独NGF刺激对DCs分泌IL-6,IL-10和IL-12无影响(均P0.05)。结论:NGF可以诱导小鼠骨髓细胞向CD11c+DCs的定向分化,并且以CD8a-亚型为主;NGF能增强LPS诱导的DC分泌IL-6,IL-10和IL-12。
[Abstract]:Aim: to investigate the role of nerve growth factor (nerve growth) in the differentiation of dendritic cells derived from bone marrow in vitro. Methods: bone marrow cells of femur and tibia were taken from C57BL / 6 mice. According to the intervention factors during the culture process, they were divided into three groups: phosphate buffer (phosphate buff ered Saline PBS) stimulation group (PBS group) and NGF-stimulated group (NGF group), granulocyte macrophage colony-stimulating factor (granulocyte monocyte colony stimulating factor-GM-CSF group and IL-4 co-stimulation group (GM-CSF IL-4 group, nn6 group), NGF plus GM-CSF and IL-4 co-stimulation group (NGF + GM-CSF and IL-4 co-stimulation group). (NGF GM-CSF IL-4 group). On the 7th day, the positive rate of CD11c cells and the ratio of CD8a- subtype were compared by flow cytometry. Immature DCswere cultured with GM-CSF IL-4 and purified by CD11c immunomagnetic bead positive sorting. The immature DCswere divided into two groups: PBS group (nong6), lipopolysaccharide polysaccharide group (LPS group nn6) and LPS group (LPS group nn6). The levels of IL-6 IL-10 and IL-12 in supernatant were detected by (enzyme linked immunosorbent Elisa. Results the percentage of CD11c DCs in NGF group was higher than that in PBS group and lower than that in GM-CSF IL-4 group (P0.05). There was no significant difference (P0.05), and in the four groups, CD8a-DCs were the main group. NGF could enhance the secretion of IL-6 IL-10 and IL-12 by LPS induced DCs (P0.05), but NGF alone had no effect on the secretion of IL-6 IL-10 and IL-12 by DCs (P0.05). Conclusion the differentiation of bone marrow cells into CD11c DCs can be induced by 1: 10% NGF, and CD8a- subtype can enhance the secretion of IL-6 IL-10 and IL-12 by LPS-induced DCs.
【作者单位】：中南大学湘雅医院呼吸内科;
【基金】：国家自然科学基金(81170023)~~
【分类号】：R329.2

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