骨髓间充质干细胞对慢性特发性血小板减少性紫癜患者T细胞T-bet和GATA-3表达的影响

摘要

目的：探讨正常人骨髓间充质干细胞体外对慢性免疫性血小板减少症患者 T 细胞 T-bet、GATA-3以及下游细胞因子 IFN-Y 和 IL-4表达的影响。方法采用 Ficoll 分离和直接贴壁、传代培养扩增骨髓间充质干细胞，用 Fi-coll 法和尼龙棉柱法获取慢性免疫性血小板减少症患者外周血的 T 淋巴细胞；以5×104个/孔的 MSCs 作为基底层细胞，经植物血凝素（PHA）刺激后，与正常人骨髓间充质细胞共培养4天，收集 T 淋巴细胞和培养上清，采用 RT-PCR 法检测 T 细胞中 T-bet 和 GATA mRNA 的表达水平，用酶联免疫吸附法（ELISA）检测培养上清干扰素- Y （IFN-Y）和白细胞介素-4（IL-4）的浓度；对照组采用地塞米松治疗（5×10-5 mol/ L）。结果实验中未检测到骨髓间充质干细胞本身分泌 IFN-Y 和 IL-4；与 T 细胞＋PHA 组相比，骨髓间充质干细胞与 T 淋巴细胞共培养组在接受PHA 刺激后，IFN-Y 的表达明显降低（P ＜0．05），IL-4表达升高（P ＜0．05）；同时显著降低 T-bet（P ＜0．05）和升高GATA-3（P ＜0．05）的表达，这与地塞米松治疗组结果相似（P ＞0．05）。结论间充质干细胞可能通过调节 T 细胞 T-bet 和 GATA-3的表达对 ITP 患者发挥治疗作用。%Objective To explore the effect of normal human bone marrow mesenchymal stem cells(MSCs)on T cell function in patients with Chronic idiopathic thrombocytopenic purpura(cITP)in vitro.Methods Normal MSCs were i-solated and cultured for proliferating to passage cells.T cells of ITP patients were harvested by using Ficoll Hypaque and nylon cotton column.MSCs of the concentrations (5X104 per well)were added to PHA induction T cell cultures with cITP patients.Then,the supernatant were respectively collected on 4 days after co-culture.The expression of IFN-γand IL-4 were studied by ELISA,and the expression of T-box (T-bet)and GATA-binding protein 3(GATA-3)were determined by RT-PCR.The control group was treated with dexamethasone (5×10-5 mol/L).Results MSCs signifi-cantly inhibit PHA induced IFN-γand T-bet expression patients with cITP(P <0.01),and increased the expression of IL-4 and GATA-3(P <0.01).Dexamethasone share the same role in the function of T,cell(P >0.05).Conclusion Human bone marrow MSCs regulates the expression of T-bet and GATA-3 and may have therapeutic implications.