活化T细胞及IFN-γ对间充质干细胞中血红素加氧酶-1抗凋亡作用的影响.docVIP

活化T细胞及IFN-γ对间充质干细胞中血红素加氧酶-1抗凋亡作用的影响 张 玲，白 海，王存邦，欧剑锋，杨国嵘 （兰州军区兰州总医院血液科，兰州 730050） 摘 要：目的 探讨免疫活性细胞和免疫调节因子对间充质干细胞（mesenchymal stem cells，MSC）及MSC中血红素加氧酶-1(HO-1)的影响。方法 体外培养人骨髓间充质干细胞, 在IFN-γ及PHA活化的T细胞刺激下, 应用RT-PCR方法检测HO-1 mRNA的表达,应用流式细胞术分析细胞凋亡情况。结果 正常骨髓间充质干细胞有HO-1 mRNA的表达，而在IFN-γ及活化的T细胞刺激后HO-1 mRNA表达下调。流式细胞术结果显示，在IFN-γ、锌原卟啉-Ⅸ(Znpp-Ⅸ)、IFN-γ+Znpp-Ⅸ的刺激下，间充质干细胞出现明显的凋亡，凋亡率分别为（56.50±0.16）%、(56.85±2.27)%、(82.53±2.65) %，较正常MSC组显著增高[（7.56±1.43）%，P 0.05]。结论 免疫活性细胞及免疫调节因子使骨髓间充质干细胞中HO-1 mRNA表达下调，进而引起MSC凋亡增加。 关键词：间充质干细胞；血红素加氧酶-1；细胞凋亡；RT-PCR；IFN-γ；Znpp-Ⅸ 中图法分类号： 文献标识码：A The influence of immunocompetent cells and immune regulator acting on heme oxygenase-1 ZHANG Ling, BAI Hai，WANG Cun-bang, OU Jian-feng, YANG Guo-rong （Department of hematology, Lanzhou General Hospital, Lanzhou 730050, China） Abstract： Objective To investigate the influence of immunocompetent cells and immune regulator act on human mesenchymal stem cells and heme oxygenase-1. Methods Human mesenchymal stem cells were cultured in vitro . RT-PCR method was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis with the stimulation of IFN-γ and T cells activated by PHA. Results The mRNA of Heme oxygenase-1 was expressed in mesenchymal stem cells and descended with the stimulation of IFN-γ and activated T cells .Adding IFN-γ and znpp, The mesenchymal stem cells apoptosis increased .While both of them added,The MSCs apoptosis was more obvious. Conclusion Immunocompetent cells and immune regulator made the expression of HO-1 mRNA in mesenchymal stem cells descending and increasing.The experimental results provided a new theoretical basis to study the pathogenesis of aplastic anemia. Key words: mesenchymal stem cells；apoptosis；heme oxygenase -1；RT-PCR；IFN-γ；Znpp-Ⅸ 骨髓间充质干细胞（mesenchymal stem cells，MSC）是一类来源于中胚层的未分化间质干细胞，是骨髓基质细胞的重要组成部分，参与骨髓造血微环境对造血的调控作用[1]。MSC具有抑制免疫的功能，而且Chabannes等[2]的研究表明，血红素加氧酶-1(heme oxygenase,HO-1)在MSC免疫抑制效应中发挥了重要作用。然而免疫细胞、免疫调节因子是如何影响MSC的，HO-1是否