地拉罗司对小鼠前成骨细胞MC3 T3－E1生物学活性的影响

摘要

目的：探讨铁鳌合剂地拉罗司（ deferasirox， DFS）体外对成骨细胞增生、分化、矿化和细胞内铁离子的影响。方法小鼠前成骨样细胞MC3T3－E1在37℃条件下体外培养，在10 mmol／L β－甘油磷酸和50 mg／L抗坏血酸的诱导作用下，分化为成骨细胞，同时用不同浓度（10、20、40μmol／L ） DFS 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， Von kos－sa染色法行细胞钙结节染色，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）、10、20、40μmol／L组A值分别为1．41±0．09、1．41±0．09、1．01±0．01、0．79±0．04、0．67±0．04； ALP 活性检测结果显示，对照组（0μmol／L）、10、20、40μmol／L组ALP活性值分别为0．73±0．03、0．65±0．02、0．54±0．03、0．35±0．04；钙结节检测结果显示，对照组（0μmol／L）、10、20、40μmol／L组TfR mRNA钙化面积比值分别为4．22±0．12、3．29±0．14、1．40±0．20、0．86±0．21； TfR mRNA表达检测结果显示，对照组（0μmol／L）、10、20、40μmol／L组TfR mRNA表达比分别为1、1．52±0．23、1．91±0．17、2．98±0．14。 MC3T3－E1细胞的增生、ALP活性、钙结节和钙化面积随DFS干预浓度的增加呈剂量依赖性降低（ P＜0．05）， TfR mRNA的表达随DFS干预浓度的增加呈剂量依赖性升高（P＜0．05）。结论 DFS可能通过螯合成骨细胞内的铁离子抑制其增生、分化和矿化。%Objective To investigate the effects of deferasirox ( DFS) on proliferation, differentiation, miner-alization and intracellular iron of osteoblast in vitro.Methods Murine preosteoblast MC3T3-E1 cells were incubated in a medium supplemented with different concentrations (10, 20, 40 μmol/L) of DFS under induction of of 10 mmol/L β-glycerophosphate and 50 mg/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.Mineralization was evaluated by von-kossa staining as-say.Transferrin receptor (TfR) mRNA was detected by real-time PCR.Results The proliferation in MC3T3-E1 cells of DMSO, control, 10, 20, 40μmol/L groups were 1.41 ±0.09, 1.41 ±0.09, 1.01 ±0.01, 0.79 ±0.04, and 0.67 ± 0.04, respectively; the ALP activity of control, 10, 20, 40 μmol/L groups were 0.73 ±0.03, 0.65 ±0.02, 0.54 ± 0.03, and 0.35 ±0.04, respectively; the minerlalized surface area and number of mineralized nodules of control, 10, 20, 40 μmol/L groups were 4.22 ±0.12, 3.29 ±0.14, 1.40 ±0.20, and 0.86 ±0.21, respectively; the expression of TfR mRNA were 1, 1.52 ±0.23, 1.91 ±0.17, and 2.98 ±0.14, respectively.Statistical analysis revealed that pro-liferation, ALP activity, minerlalized surface area and number of mineralized nodules were decreased by DFS in a con-centration-dependent manner (P<0.05), and the expression of TfR mRNA was up-regulated by DFS in a concentration-dependent manner (P<0.05).Conclusion DFS can significantly inhibit proliferation, differentiation and mineraliza-tion of osteoblasts and the mechanism behind inhibition may involve decreased levels of intracellular iron.