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。
Objective To investigate the effects of deferiprone （DFP） on proliferation, differentiation, and in- tracellular 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/L 13-glycerophosphate and 50 μg/mL 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 ceils in DMSO, control, 25, 50, 100 μmol/L of DFPgroups 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 un derlie the inhibition.
出处
《中华骨质疏松和骨矿盐疾病杂志》
CSCD
2017年第2期121-124,共4页
Chinese Journal Of Osteoporosis And Bone Mineral Research
基金
江苏省重点研发计划(社会发展)项目(BE2016720)
江苏省妇幼健康科研项目(F201504)
镇江市科技计划项目(SH2014031)
