Xie Fang, Zhao Ming-Feng, Zhu Hai-Bo, Xiao Xia, Xu Xin-Nü, Mu Juan, Li Yu-Ming
Department of Hematology and Oncology, Tianjin First Central Hospital, Tianjin 300192, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Aug;19(4):1038-42.
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 cells (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 function 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 disorders.
本研究旨在通过将骨髓单个核细胞与铁共培养建立铁过载骨髓（BM）模型，并研究其造血变化。通过向骨髓单个核细胞中添加不同浓度的柠檬酸铁（FAC）并培养不同时间来建立铁过载模型，并通过检测不稳定铁池（LIP）来确认该模型。然后测定造血细胞的凋亡、造血集落形成能力（CFU-E、BFU-E、CFU-GM和CFU-mix）以及骨髓细胞中CD34(+)细胞的百分比。在用去铁胺（DFO）处理铁过载骨髓后检测这些指标的变化。结果显示，骨髓细胞在不同浓度的FAC下培养不同时间后，LIP呈时间和浓度依赖性增加。当在400 μmol/L的FAC下培养24小时时，细胞内LIP达到最高水平。对骨髓细胞造血功能的检测发现，与正常对照（8.9 ± 0.96%）相比，FAC处理细胞的凋亡率（24.8 ± 2.99%）显著增加（p < 0.01）。与正常对照相比，FAC处理细胞的造血集落形成能力明显降低（p < 0.05）。FAC处理细胞中CD34(+)细胞的百分比（0.39 ± 0.07%）也与正常对照（0.91 ± 0.12%）相比显著降低（p < 0.01）。并且通过添加DFO可以缓解这些变化。结论是通过在体外向骨髓单个核细胞中添加铁建立了铁过载模型，铁过载骨髓的造血功能存在缺陷。通过用DFO处理从骨髓细胞中去除过量铁可以缓解这些变化。这些发现将有助于进一步研究铁过载对骨髓造血的机制，也有助于找到治疗患有造血障碍的铁过载患者的方法。
