去铁胺对高铁环境下成骨细胞分化的影响

Effect of deferoxamine on osteoblast differentiation under iron overload environment

目的观察去铁胺（DFO）对于高铁环境[利用枸橼酸铁铵（FAC）提供铁离子制作高铁环境]下成骨细胞分化的影响。方法通过细胞计数试剂盒（CCK-8）法确定FAC及DFO的干预计量。实验分为3组：对照组、50 μmol/L FAC干预组、FAC＋DFO组（50 μmol/L FAC干预同时加入20 μmol/L DFO）。干预后3组细胞分别行碱性磷酸酶（ALP）染色和活性定量，茜素红染色、钙结节计数，对成骨相关基因[Runt相关基因2（Runx2）、锌指转录因子SP7、骨γ羧基谷氨酸（Bglap）、骨保护素（OPG）]利用实时荧光定量核酸扩增检测系统（QPCR）检测其mRNA的表达水平。结果CCK-8结果显示，50 μmol/L FAC干预组成活率为（95.6±0.7）%，20 μmol/L DFO干预组成活率[（97.6±0.5）%]不影响细胞活性，各组差异无统计学意义（P＝0.427）。与对照组比较，FAC干预组，成骨相关基因Runx2、Sp7、Bglap、OPG表达下降；而FAC＋DFO干预组与FAC干预组比较成骨相关基因表达上升，差异有统计学意义（P＝0.026）。ALP染色及活性定量和茜素红染色及结节计数与对照组比较，FAC组结节计数（36±3）下降，而FAC＋DFO组结节计数（120±6）比FAC组上升，差异有统计学意义（P＝0.017）。结论在高铁环境中，成骨细胞分化受到抑制；DFO干预可在一定程度上恢复高铁环境下成骨细胞分化。

ObjectiveTo study the influence of eferoxamine （DFO） intervention on osteoblast defferentiation under iron overload environment [using ferric ammonium citrate （FAC） to offer iron overload environment].MethodsThe FAC and DFO intervention dosage was determined by method of cell counting kit-8 （CCK-8）. Experiment was divided into three groups： control group, 50 μmol/L FAC intervention group, and FAC ＋ DFO group （50 μmol/L FAC intervention ＋ 20 μmol/L DFO）. After intervention, alkaline phosphatase staining and quantitation, alizarin red staining and calcium nodule count were carried out. Real-time quantitative polymerase chain reaction detecting system （QPCR） was used to detect the osteogenesis related gene [runt related transcription factor 2 （Runx2）, special protein 7 （Sp7）, bone gamma-carboxyglutamic acid-containing protein （Bglap）, osteoprotegerin （OPG）] mRNA expression level.ResultsCCK-8 showed 50 μmol/L FAC [（95.6±0.7）%], or 20 μmol/L DFO [（97.6±0.5）%] did not influence the cell viability, with the difference being not statistical differences among groups （P=0.427）. As compared with control group, the expression of osteogenesis related genes （Runx2, Sp7, Bglap and OPG） was significantly decreased in the FAC group. As compared with the FAC group, the expression of osteogenesis related genes was significantly increased in the FAC ＋ DFO group （P=0.026）. As compared with the control group, the activity of alkaline phosphatase （ALP） staining and quantitation, alizarin red staining and nodule count were decreased in the FAC group. As compared with the FAC group （36±3）, the nodule count in the FAC ＋ DFO group （120±6） was increased significantly （P=0.017）.ConclusionOsteoblast differentiation is restrained under the iron overload environment. DFO intervention can restore osteoblast differentiation to a certain extent under the iron overload environment.