目的观察不同因素刺激下的DC分化及数量变化,对CD4+CD25+调节性T细胞的抑制作用。方法制备六味地黄丸含药血清,刺激树突状细胞;反复冻融法制备细胞抗原负载树突状细胞;将实验分为:DC与空白血清组、DC与胃癌细胞组、DC与含药血清组、DC与胃癌细胞和六味地黄丸含药血清联合作用组,将各组分别与淋巴细胞共培养,MTT法检测各组DC数量的变化,流式细胞仪检测DC对CD4+CD25+调节性T细胞的作用。结果在药物作用48 h含药血清浓度为15%时与空白血清组分别作用24、72 h相比,DC的吸光光度值能显著升高(P〈0.05)。胃癌细胞与六味地黄丸共同作用组DC吸光光度值显著升高,DC吸光光度值升高对CD4+T细胞的增殖能力有明显调节作用,可降低CD4+T细胞中CD4+CD25+调节性T细胞的数量。结论胃癌细胞作为抗原负载DC,具有促进DC细胞增殖及分化的作用,并可增强CD4+T细胞的增殖能力和降低CD4+T细胞中Treg的数量,六味地黄丸具有辅助调节作用。
【Objective】The differentiation and changes of DC in the number of of different stimuli, inhibition of CD4+CD25+regulatory T cells.【Methods】Preparation of Six Ingredient Rehmannia Pill medicated serum, stimulation of dendritic cells; using repeated freezing and thawing method to prepare cell antigen loading dendritic cell;divided into four groups: DC with the blank serum group, DC group, DC and gastric cancer cell and medicated serum group, the combined effect of DC cells with gastric cancer and Six Ingredient Rehmannia Pill medicated serum group; each group co culture with lymphocyte respectively, changes in the number of DC was detected by MTT,using flow cytometry to detected DC and its effect on CD4+CD25+ regulatory T cells. 【Results】Effect of 48 h respectively roles in drug containing serum concentration is 15% and the blank serum group 24 h, 72 h than the DC photometric value can be increased significantly(P0.05). Gastric cancer cells and the six tastes glutinous rehmannia pill interaction group DC photometric values increased significantly, DC photometric values increased on CD4+ T cells proliferation ability has obvious adjustment function,which can reduce the number of CD4+CD25+ regulatory T cells in CD4+T cells. 【Conclusion】Gastric cancer cells as antigen loaded DC, has the function of promoting DCs proliferation and differentiation, and can enhance the CD4+T cell proliferation ability and reduce the amount of Treg in CD4+ T cells; the six tastes glutinous rehmannia pill has the auxiliary regulating function.