Ginsenoside Rg3 affecting mature and function of dendritic cells
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摘要: 目的 探讨人参皂苷单体Rg3对外周血来源的树突状细胞(dendritic cell,DC)成熟及其功能的影响。 方法 ①在GM-CSF和IL-4联合作用下,外周血单个核细胞诱导分化为未成熟DC(imDC),第5天加入TNF-α促进成熟,并加入不同剂量的Rg3,分别设为低、中、高剂量组(Rg3浓度分别为10、20、40 μg/ml)及空白对照组,诱导出Rg3-DC。流式细胞术检测诱导第7、14天细胞表面CD80、CD86及CD83的表达;②收集培养第7、14天时各组Rg3-DC,进行混合淋巴细胞反应,采用MTT法测定Rg3-DC刺激异体淋巴细胞增殖指数;③ELISA方法检测各剂量组Rg3-DC分泌IL-12的水平。 结果 ①人参皂苷Rg3中、高剂量组培养第7、14天后的DC表面标志CD80、CD86及CD83表达明显增高,与对照组比较差异有统计学意义,其中DC成熟标记CD83在中、高剂量组培养14 d后分别为(38.6±2.21)%、(44.21±4.94)%,与对照组比较差异均有统计学意义(P<0.05及P<0.01)。②高剂量组Rg3-DC刺激异体淋巴细胞增殖指数高于其他3组(P<0.05)。③高剂量组Rg3-DC3-DC分泌IL-12浓度为(34.45±4.42)pg/ml,高于对照组的(10.24±8.82)pg/ml(P<0.05)。 结论 人参皂苷Rg3可诱导DC成熟,促进其功能转变,在一定浓度下呈剂量依赖性。Abstract: Objective To investigate the effect of ginsenoside Rg3 on the maturation and function ofhuman peripheral blood monocyte-derived dendritic cells(DC). Methods ①The differentiation of peripheral blood mononuclear cells(PBMC) to immature DC(imDC) was induced by the combination of GM-CSF and IL-4.On the fifth day,TNF-α was added to make the mature of DC,so as different doses of Rg3,which were divided into low dose group(10 μg/ml),middle dose group(20 μg/ml),high dose group(40 μg/ml) and control group.The expression of CD80,CD86 and CD83 on DC cell surface which induced by Rg3 at the 7th and 14th day were detected byflow cytometry.②Rg3-DCs were collected at the 7th and 14th day,then mixed lymphocyte reaction was performed,MTT method was used to analyze the influence of ginsenoside Rg3 and DCs on the stimulation of allogeneic lymphocyte proliferation;③The interleukin-12(IL-12) level produced by Rg3-DCs were detected with ELISA. Results ①The expression of DC markers CD80,CD86 and CD83 on cell surface of middle and high Rg3 dose groups were significantly increased after culture for 7 and 14 days,the difference was significant as compared with the control group(P<0.05).The mature markers of CD83 at the 14th day of middle and high dose groups were (38.60±2.21)% and (44.21±4.94)%,respectively,which was significantly higher than that of the control group (28.45±7.20)%,P<0.05 and P<0.01.①Rg3-DCs showed more effective in the high dose group on stimulating lymphocyte proliferation when compared with the other groups(P<0.05).③In the high dose group,IL-12 level produced by Rg3-DC was (34.45±4.42)pg/ml,which was higher than that of the control group (10.24±8.82)pg/ml,P<0.05. Conclusion Ginsenoside Rg3 can induce DC maturation,promote thefunctional conversion.A dose-dependent can be observed within a certain concentration.
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