【摘要】 目的研究铁螯合剂去铁酮(deferiprone,DFP)体外对成骨细胞增生、分化以及细胞铁代谢的影响。方法体外培养小鼠前成骨样细胞MC3T3-E1,在10 mmo L/Lβ-甘油磷酸和50μg/m L抗坏血酸的诱导下,分化为成骨细胞,同时用不同浓度(25、50、100μmol/L)DFP干预,用CCK-8法检测细胞的增生,碱性磷酸酶(alkaline phosphatase,ALP)活性试剂盒检测细胞ALP活性,实时定量PCR检测细胞膜转铁蛋白受体(transferrin receptor,TfR)mRNA的表达。结果 MC3T3-E1细胞增生结果显示,DMSO溶剂组、对照组(0μmol/L)和DFP 25、50、100μmol/L组A值分别为1.36±1.10、1.41±0.09、0.78±0.06、0.68±0.03、0.46±0.01;ALP活性检测结果显示,对照组(0μmol/L)和DFP 25、50、100μmol/L组ALP活性值分别为0.83±0.05、0.75±0.04、0.64±0.03、0.51±0.03;TfR mRNA表达检测结果显示,对照组(0μmol/L)和DFP 25、50、100μmol/L组表达比分别为1、1.16±0.05、1.32±0.06、2.30±0.11。MC3T3-E1细胞的增生、ALP活性随DFP干预浓度的增加呈剂量依赖性下降(P<0.05),TfR mRNA的表达随DFP干预浓度的增加呈剂量依赖性上升(P<0.05)。结论 DFP可能通过螯合成骨细胞内的铁离子抑制其增生、分化。更多还原

【Abstract】 Objective To investigate the effects of deferiprone(DFP) on proliferation,differentiation,and intracellular iron metabolism of preosteoblasts in vitro. Methods Murine preosteoblasts of MC3T3-E1 were incubated in a medium supplemented with different concentrations(10,25,100 μmol/L) of DFP under induction of 10 mmol/Lβ-glycerophosphate and 50 μg/m L L-ascorbic acid. Proliferation of MC3T3-E1 cells was evaluated by CCK-8 assay. Alkaline phosphatase(ALP) activity was measured using ALP viability kit. Transferrin receptor(TfR) mRNA was detected by real-time PCR. Results The proliferation indices of MC3T3-E1 cells in DMSO,control,25,50,100 μmol/L of DFP groups were 1. 36 ± 1. 10,1. 41 ± 0. 09,0. 78 ± 0. 06,0. 68 ± 0. 03,and 0. 46 ± 0. 01,respectively. The ALP activity of control,25,50,100 μmol/L of DFP groups were 0. 83 ± 0. 05,0. 75 ± 0. 04,0. 64 ± 0. 03,and 0. 51 ±0. 03,respectively. The relative mRNA expression of TfR were 1,1. 16 ± 0. 05,1. 32 ± 0. 06,and 2. 30 ± 0. 11,respectively. The proliferation and ALP activity were decreased by DFP in a concentration-dependent manner(P < 0. 05),and the mRNA expression of TfR was up-regulated by DFP in a concentration-dependent manner(P < 0. 05). Conclusion DFP can significantly inhibit proliferation and differentiation of osteoblasts,decreased levels of intracellular iron may underlie the inhibition.更多还原