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| 铁调素干预成骨细胞(hFOB1.19)后细胞膜转铁蛋白1、铁离子、矿化及成骨基因变化研究 |
| The changes of ferroportin 1, iron ion, and mineralization and ossific genes in hFOB 1.19 cells after the intervention with hepcidin |
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| DOI：10.3969/j.issn.1006-7108.2013.10.001 |
| 中文关键词: 铁调素 铁离子 膜转铁蛋白1 细胞矿化 成骨基因 |
| 英文关键词:Hepcidin Iron ion Ferroportin1 Mineralization Ossific gene |
| 基金项目:国家自然科学基金项目（81273090）；江苏省自然科学基金项目（BK2012608）；苏州大学青年预
研基金（SDY2011A42）；苏州大学附属第二医院青年职工预研基金（SDFEYQN1105） |
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| 中文摘要: |
| 目的 探讨铁调素对成骨细胞铁代谢指标、骨代谢指标的影响和相互关系。方法 成骨细胞
（hFOB1.19）培养3d后，使用逆转录-聚合酶链反应（RT-PCR）检测成骨细胞（hFOB1.19）的膜转铁蛋白1
（ferroportin1，Fpn1）的表达；再用不同浓度的铁调素（25noml/L，50noml/L，100noml/L）干预培养
环境，对照组加相同体积双蒸水，干预20h，MTT法检测细胞增殖情况；激光共聚焦扫描显微镜（CLSM）检
测成骨细胞内铁离子荧光染色后荧光强度变化情况；Von Kossa染色检测成骨细胞矿化情况；RT-PCR检测
骨钙素（BGP）、I型胶原（COL1）、骨保护素（OPG）基因表达。结果 1、成骨细胞存在Fpn1表达；2、铁
调素对成骨细胞增殖无显著影响（P>0.05）；3、铁调素干预后，成骨细胞内铁离子含量增多，且与铁调
素干预存剂量依赖性关系（P<0.05）；4、铁调素干预后，成骨细胞矿化功能增强，且与铁调素干预存剂
量依赖性关系（P<0.05）；5、RT-PCR检测显示不同浓度铁调素干预后，各组COL1、OPG、BGP mRNA均有表
达；不同浓度组的COL1、OPG、BGP mRNA表达光密度比值不同，组间密度比值比较存在统计学意义
（P<0.05），但铁调素在400nmol/L时抑制OPG的表达（P<0.05）。结论 成骨细胞存在铁调素作用的膜转
铁蛋白-1，所以铁调素干预成骨细胞培养环境后细胞内铁离子发生特异性变化，同时细胞矿化指标显著增
加、细胞COL1、OPG、BGP mRNA表达含量增加；研究提示：铁调素可能通过膜转铁蛋白影响成骨细胞铁离
子变化，细胞铁代谢变化可能引起细胞成骨指标上调改变。 |
| 英文摘要: |
| Objective To investigate the effect of hepcidin on iron and bone metabolism
indexes in osteoblasts and their correlation. Methods After 3-day culturing, the expression
of ferroportin1 (Fpn1) in human fetal osteoblasts hFOB1.19 was detected using reverse
transcriptase-polymerase chain reaction (RT-PCR). Then, different concentrations of hepcidin
(25noml/L, 50noml/L, and 100noml/L) were additioned to the medium, and the same volume of
double distilled water was additioned in control group. After 20-hour intervention, cell
proliferation was detected using MTT method. The changes of fluorescence intensity after
fluorescence staining of intracellular iron in osteoblasts were measured using a confocal
laser scanning microscope (CLSM). von Kossa staining was performed to evaluate cell
mineralization. The expression of BGP, COL1, and OPG was detected using RT-PCR. Results The
expression of Fpn-1 was positive in hFOB 1.19. Hepcidin had no significant effect on hFOB
1.19 proliferation (P>0.05). After the intervention with hepcidin, the concentration of ion
increased in osteoblasts, and the mineralization also enhanced, both showing a dose-
dependent manner with the concentration of hepcidin (P<0.05). After the intervention with
different concentrations of hepcidin, the mRNA expression of COL1, OPG, and BGP detected
using RT-PCR was positive in all groups. The optical density ratio of COL1, OPG, and BGP
after the intervention with different concentrations of hepcidin was different, and the
difference was significant (P<0.05). The expression of OPG was inhibited by hepcidin at the
concentration of 400 nmol/L (P<0.05). Conclusion Fpn1, which can react with hepcidin, exsits
in osteoblasts. So, after the intervention of hepcidin, the intracellular concentration of
iron ion changes specificly. And the mineralization enhances. The mRNA expression of BGP,
COL1, and OPG also increase. Hepcidin may affect the changes of iron ion concentration
through Fpn1 in osteoblasts, and the changes of iron metabolism may result in the up-
regulation of osteogenesis indexes. |
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