铁过载骨髓造血细胞体外模型的建立及其对造血的影响  

Establishment of Iron Overloaded Bone Marrow Model In Vitro and Its Impact on Hematopoiesis

作　　者：谢芳[1] 赵明峰[1] 朱海波[1] 肖霞[1] 徐新女[2] 穆娟[1] 李玉明[1] 

机构地区：[1]天津医科大学第一中心临床学院,天津市第一中心医院血液科,天津300192 [2]天津医科大学第一中心临床学院,国家卫生部急救医学重点实验室,天津300192

出　　处：《中国实验血液学杂志》2011年第4期1038-1042,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金(编号81041043);教育部留学归国人员科研启动基金(编号教外司留[2007]1108);天津市卫生局科技基金(编号05KY10)

摘　　要：本研究通过铁与骨髓单个核细胞共培养,建立铁过载骨髓造血细胞模型并观察其对造血功能的影响。在体外培养骨髓单个核细胞的过程中,在培养液中添加枸橼酸铁铵(FAC),使细胞铁过载,建立铁过载骨髓造血细胞模型,然后检验这一过程中造血细胞的凋亡水平、造血集落形成(CFU-E、CFU-GM、BFU-E和CFU-mix)和CD34+细胞的计数变化。再用去铁胺(DFO)祛铁,观察上述指标的变化。结果发现:在不同时间加入培养液中不同浓度FAC,骨髓造血细胞内可变铁池(LIP)水平升高,且具有时间和浓度依赖性,在含400μmol/L FAC的培养液中培养24小时时LIP水平达到最高。骨髓细胞造血功能检测表明,FAC组细胞凋亡比例(24.8±2.99%)较对照组(8.9±0.96%)显著升高(p(0.01);造血集落形成单位计数明显低于对照组(p(0.05);CD34+细胞比例(0.39±0.07%)与对照组(0.91±0.12%)相比也显著降低(p(0.01)。这些损伤影响可以通过DFO的处理部分消除。结论:在体外培养过程中添加铁能诱导骨髓单个核细胞铁过载,建立铁过载骨髓造血细胞模型,并能引起骨髓造血功能损伤,而这种损伤作用可以通过祛铁法减轻。本模型为深入研究铁过载对骨髓细胞造血功能的作用机制提供实验方法,并为骨髓造血功能低下的铁过载病人治疗提供新的靶点。This study was to establish an iron overload bone marrow （BM） model by co-culturing the mononuclear cells from BM with iron, and investigate its hematopoiesis changes. The iron overload model was set up by adding different concentration of ferric citrate （FAC） into the mononuclear cells from BM and culturing for different time, and the model was confirmed by detecting labile iron pool （LIP）. Then the apoptosis of hematopoietic cells, ability of hematopoietic colony forming （ CFU-E, BFU-E, CFU-GM and CFU-mix） and percentage of the CD34± cells of the BM cells all were determined. The changes of these indexes were tested after the iron-overloaded BM was treated with deferasirox （DFO）. The results showed that after BM cells were cultured with FAC at different concentrations for different time, the LIP increased in time-and concentration-dependent manners. The intracellular LIP reached maximum level when cultured at 400 μmol/L of FAC for 24 hours. The detection of BM cell hematopoietic function found that the apoptotic rate of the FAC-treated ceils （24.8 ± 2.99% ） increased significantly, as compared with normal control （8.9 ± 0.96% ） （p 〈 0.01 ）. The ability of hematopoietic colony forming in FAC-treated cells decreased markedly, as compared with normal control （p 〈0.05）. The percentage of CD34＋ cells of FAC-treated cells （0.39 ±0.07% ） also decreased significantly, as compared with normal control （0.91 ± 0.12% ）（p 〈 0.01 ）. And these changes could be alleviated by adding DFO. It is concluded that the iron-overloaded model has been set by adding iron into the mononuclear cells from BM in vitro, and the hematopoietic funtion of iron-overloaded BM is deficient. These changes can be alleviated by removing the excess iron from the BM cells through treating with DFO. These findings would be helpful to further study the mechanism of iron-overload on the hematopoiesis of BM and also useful to find the way to treat iron-overload patients with hematopoietic disorde

关 键 词：铁过载 骨髓细胞模型 造血功能 可变铁池 

分 类 号：R331.22[医药卫生—人体生理学]