绿原胶囊浸出液促成人外周血树突状细胞体外成熟的研究  

Research of Luyuan Capsule Leachate on the in Vitro Maturation of Dendritic Cells of Adult Peripheral Blood

作　　者：龙丹[1] 王佳[2] 李典[3] 李胜富[1] 

机构地区：[1]四川大学华西医院卫生部移植工程与移植免疫重点实验室,成都610041 [2]四川大学华西医院风湿免疫科,成都610041 [3]四川大学华西医院药剂科,成都610041

出　　处：《中国中西医结合杂志》2012年第11期1558-1561,共4页Chinese Journal of Integrated Traditional and Western Medicine

摘　　要：目的探讨绿原胶囊浸出液对成人外周血来源的树突状细胞(Dendritic cells,DCs)增殖及成熟的影响。方法采集健康男性(4名)外周血各100mL,采用密度梯度离心法获取外周血单个核细胞,贴壁法获取弱贴壁的DCs前体细胞,用添加重组人粒细胞-巨噬细胞集落刺激因子(recombinant human GM-CSF,rhGM-CSF)和重组人白细胞介素4(recombinant human IL-4,rhIL-4)的RPML-1640的DCs培养基培养DCs前体细胞7天,可获得未成熟的DCs。以在DCs培养体系中加入1%浓度的绿原胶囊浸出液共培养未成熟DCs2天作为实验组,以20ng/mL重组人肿瘤坏死因子(Recombinant human TNF-α,rh TNF-α)共培养未成熟DCs2天作为对照组。倒置相差显微镜观察细胞形态,细胞计数法检测成熟DCs增殖能力,流式细胞仪检测成熟DCs表达的特异性标记物CD83。结果与对照组相比,实验组培养2天后,DCs的增殖能力提高,DCs表面呈现出比对照组更多和更长的树突状突起,实验组CD83表达[(84.1±0.3)%]明显高于对照组[(58.7±0.2)%],两组比较差异有统计学意义(P<0.05)。结论 1%浓度的绿原胶囊浸出液可促进外周血来源的DCs成熟和增殖,且作用较20ng/mL的rh TNF-α更强。Objective To investigate the effect of Luyuan Capsule （LYC） leachate on the proliferation and maturation of dendritic cells （DCs） of adult peripheral blood. Methods Totally 100 mL peripheral blood was collected from 4 healthy males. The peripheral blood mononuclear cells were isolated using density gradient centrifugation. The precursor DCs were obtained using wall adherent culture. The immature DCs with recombinant human GM-CSF （rhGM-CSF） and recombinant human IL- 4 （rhIL- 4） added were harvested for 7 days to get immature DCs. The immature DCs cultured with 1% LYC leachate for two days were taken as the experimental group, while the immature DCs cultured with 20 ng/mL recombinant human TNF-α （rhTNF-α） for two days were taken as the control group. The morphology of DCs was observed by inverted phase contrast microscope, and the cell proliferation was detected by cell counting. The DCs specific marker CD83 was analyzed using flow cytometry. ResultsThe proliferation capacities of DCs were improved in the experimental group. More and longer dendritic protrusions and possess were shown on the surface of DCs in the experimental group than in the control group. The CD83 expressions was obviously higher in the experimental group （84.1%±0.3%） than in the control group （58.7%±0.2%）, showing statistical difference （P0.05）. Conclusion 1% LYC leachate could promote the maturation and proliferation of peripheral blood DCs, stronger than that of 20 ng/mL rhTNF-α.

关 键 词：绿原胶囊 树突状细胞 成熟 增殖 

分 类 号：R967[医药卫生—药理学]