摘要
目的探讨黄芪多糖对钙离子载体A23187快速诱导树突状细胞（DC）免疫活性的影响。方法2014年9月20日，无菌条件下分别于9例健康男性志愿者肘静脉采集血液10～20mL，作为实验材料。志愿者纳入标准：采血当天无发热、无感染，无肝炎、结核等传染病接触史。采用Ficoll密度梯度离心法分离外周血单个核细胞（PBMC），取生长状态良好的PBMC（浓度为1．0×10^6／mL），以1mL／孔接种于24孔平底培养板。选择9个培养孔，按简单随机法将其分为3组，分别加入不同的诱导剂。A23187＋黄芪多糖组（n=3）：先加入A23187，再加入黄芪多糖；A23187组（n=3）：仅加入A23187；对照组（n=3）：不加诱导剂。3组细胞均于37℃、5％CO2及饱和湿度培养箱中培养48h后，比较各组细胞形态学、免疫表型及刺激同种异体T淋巴细胞增殖能力的差异。结果体外培养48h后，A23187组部分PMBC出现树状突起，DC特异性抗原CD83、CD80、CD86分子表达阳性率分别为（28．9±6．2）％、（42．1±5．6）％及（49．1±7．8）％；A23187＋黄芪多糖组的绝大部分PMBC出现明显树突状突起、且免疫表型及刺激同种异体T淋巴细胞增殖的能力均较A23187组PBMC增强。A23187＋黄芪多糖组细胞表面DC特异性抗原CD83、CD80、CD86分子表达阳性率分别为（43．6±8．2）％、（69．9±19．7）％及（79．4±22．5）％，与A23187组及对照组比较，差异均有统计学意义（t=-65．23、-144．86、-81．28，P〈0．05；t=-25．50，-82．36，-45．21；P〈0．05）。在1：1280、1：320、1：80、1：20、1：5的不同刺激细胞（SC）与效应细胞（EC）比例下，A23187＋黄芪多糖组DC细胞刺激指数（SI）分别为1．38±0．39、3．91±0．94、7．91±1．48、10．38±2．46、11．7±2．83。与A223187组及对照组相比，差异有统计学意义（t=-32．45、-65．96、-45．32、-37．89、-64．25，P〈0．05；t=-5�
Objective To investigate the effect of astragalus polysaccharides on the immune activity of dendritic cells （DC） induced by calcium ionophore A23187. Methods On 20th September 2014, 10- 20 mL whole blood were collected from 9 healthy male volunteers through cubital vein under aseptic condition, respectively. Volunteers inclusion criteria： no fever, no infection, and no history of exposure to hepatitis, tuberculosis and other infectious diseases. Using Ficoll density gradient centrifugation to obtain peripheral blood mononuclear cells （PBMC）, and selected PBMC in good growth conditions （concentration is 1.0×10^6/mL） to culture in the flat culture plate of 24 holes with 1 mL per hole. Nine holes of these culture plate were divided into 3 groups by the simple random method, and each group was added different inducing agent as follow： A23187＋astragalus polysaccharides group （n=3）： added A23187 and astragalus polyose; sequentially; A23187 group （n=3）： added A23187 only; control group （n=3）： did not add any inducing agent. Cells of these 3 groups are all cultivated in 37℃, 5% CO2 and saturated humidity incubator for 48 h, and then compared the changes of cytomorphology, expressions of immunophenotype and ability of stimulating proliferation of allogeneic T lymphocytes, respectively. Results After cultured for 48 h, the changes of morphological, expressions of immunophenotype and ability to stimulating proliferation of allogeneic T lymphocytes were enhanced of PBMC in A23187＋astragalus polysaccharides group compared to those of A23187 group and control group. After cultured for 48 h, some of the PMBC appeared dendritic processes in A23187 group, and vast majority of PBMC showed obvious dendritic processes in A23187＋ astragalus polysaccharides group. The positive rate of CD83 ,CDS0 and CD86 expressing on DC in A23187＋ astragalus polysaccharides group increased significantly compared to those of A23187 group[（43.6±8.2）% vs （28.9±6.2）%, t=-25.50, P〈0.05; （69.9±19.
出处
《国际输血及血液学杂志》
CAS
2016年第2期114-119,共6页
International Journal of Blood Transfusion and Hematology
基金
四川省科技厅基金（2012JY0042）
关键词
树突状细胞
黄芪多糖
钙离子载体
中草药
Dendritic cells, Astragalus polysaccharides
Calcium ionophore
Drugs, Chinese herbal
