目的:本研究试图检测不同浓度17β雌二醇(E2)存在的情况下人胎肺间充质干细胞(MSC)增殖和表型的变化,并分析这种状态的MSC对树突状细胞(DC)成熟和功能影响及其可能的机制。方法:分离培养人胎肺MSC,加入不同浓度F2,在24小时后对MSC进行细胞计数、测定其增殖和贴壁能力,并用流式细胞仪分析MSC表面标志,RT-PCR测定MSC中细胞因子(IL-6、TGF-β和VEGF)mRNA的表达情况。进一步探讨了E2处理24小时后的MSC对Dc成熟和功能的影响。结果:分离得到的MSC纯度达到95%以上;E2影响MSC的增殖和贴壁能力,但不影响MSC表面标记的表达;MSC与DC共培养后DC表面CD86、MHCⅡ和CD80的表达均有所降低,而当DC与经E2预处理过的MSC共培后,DC表面MHCH、CD80和CD86的表达回升;高浓度E2作用MSC24小时后,MSC表达的TGF-β与对照组相比减少,而IL-6和VEGF与对照组相比增加。结论:E2可能通过调节MSC分泌细胞因子的水平,改变MSC对DC的免疫抑制作用。
Objective:To investigate the effects of 17β-Estradiol (E2) on mesenchymal stem cells (MSC) and to evaluate the'effects of MSC treated with E2 on the maturation and function of dendritic cells (DC). Methods: We first isolated and cultured MSC from the human fetal lung. The MSC were treated with E2 for 24 hours at various concentrations (10-9,10-s and 10-7 tool/L). After cell counting, proliferation, adherent ability and immunophenotypes of MSC were detected by flowcytometry. The gene expressions of cytokine (IL-6, TGF-β and VEGF) were measured by RT-PCR.The effects of MSC treated with E2 on the maturation and function of DC were determined. Results: After treated with E2, the proliferation and adherent ability of MSC were increased, while the immunophenotypes of MSC were not affected. When MSCs co-cultured with DC, MSC could inhibit the immuophenotypes and function of DC. However, when DC co-cultured with E2-pretreated MSC, the immunophenotypes (MHCⅡ, CDSO and CD86) of DC had been reconstructed. After treated with the high concentration of E2 for 24 hours, MSC secreted lower level of TGF-β than that in the control group, while IL-6 and VEGF expressions were increased compared with those in the control group. Conclusion: Estrogen may alter the immuno-suppressive effects of MSC on DC via modulating the cytokine secretion of MSC.