中国骨质疏松杂志

高铁培养环境对小鼠前成骨细胞MC3T3-E1增殖、分化的影响

Effect of excess iron culturing on the proliferation and differentiation of mouse MC3T3-E1 preosteoblasts

DOI：10.3969/j.issn.1006-7108.2013.10.003

英文关键词:Iron ion Osteoblast Differentiation

基金项目:国家自然科学基金(81273090)；江苏省自然科学基金(BK2012608)；苏州市科技基础设施建设计划之高技术研究重点实验室资助项目(SZS2012208)；苏州市应用基础研究计划之医疗卫生部分资助项目 (SYS201002)

作者	单位
赵国阳1 何银锋2 李光飞2 Xi Hang3 徐又佳2	1. 苏州大学附属医院骨一科，镇江 212001； 2. 苏州大学附属第二医院骨科，苏州215004； 3.美国纽约大学医学院，美国纽约

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中文摘要:

目的观察高铁培养环境下小鼠前成骨样细胞MC3T3-E1增殖、分化指标的变化趋势，探讨铁离子对成骨细胞增殖、分化的影响。方法小鼠前成骨样细胞MC3T3-E1在37℃条件下体外培养，在10mmol/Lβ-甘油磷酸和50μg /mL抗坏血酸的诱导分化的作用下，分化为成骨细胞，同时用不同浓度(50、100、 200μmol/L)枸橼酸铁铵(FAC)干预，用MTT法检测细胞的增殖活性，RT-PCR法检测成骨细胞分化基因成骨相关转录因子(Runx2) 、锌指结构转录因子(Osterix) 、骨唾液酸蛋白(BSP)和骨钙素(OC)的表达，碱性磷酸酶(ALP)活性试剂盒检测细胞碱性磷酸酶活性。结果 MC3T3-E1细胞的增殖活性、成骨分化相关基因的表达以及ALP水平随FAC干预浓度的增加呈剂量依赖性降低(P<0.05)。结论高铁培养环境可明显抑制小鼠前成骨样细胞MC3T3-E1的增殖和分化。

英文摘要:

Objective To observe the changing pattern of proliferative and differentiatial indexes of mouse MC3T3-E1 preosteoblasts in the culture with excess iron, and to investigate the effect of iron on the proliferation and differentiation of osteoblasts. Methods Mouse MC3T3-E1 preosteoblasts were cultured at 37oC in vitro. Under the induction of 10mmol/l β- glycerophosphate and 50μg/ml L-ascorbic acid, MC3T3-E1 preosteoblasts differentiated into osteoblasts. Meanwhile, different concentrations (50, 100, and 200 μmol/l) of FAC were additioned into the medium for intervention. Proliferation of MC3T3-E1 cells was detected using MTT method. The mRNA expressions of Runx2, Osterix, bone sialoprotein (BSP), and osteocalcin (OC) were detected using RT-PCR. ALP activity was measured using ALP viability kit. Results The proliferation of MC3T3-E1 cells, the mRNA expressions of Runx2, Osterix, BSP, and OC, and ALP activity decreased along with the increase of FAC concentration, showing a dose-dependent manner (p<0.05). Conclusion Excess iron culturing can significantly inhibit the proliferation and differentiation of mouse MC3T3-E1 preosteoblasts.

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