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Ficoll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RTPCR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Tbet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.
出处
《中国实验诊断学》
2014年第5期697-700,共4页
Chinese Journal of Laboratory Diagnosis
基金
湖北医药学院博士启动基金(2009QDJ36)
