CAJ | 학술논문

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3 d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（25 noml/L，50 noml/L，100 noml/L）干预培养环境，对照组加相同体积双蒸水，干预20 h，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400 nmol/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。
목적：탐토철조소대성골세포철대사지표、골대사지표적영향화상호관계。방법성골세포（ hFOB1.19）배양3 d후，사용역전록-취합매련반응（RT-PCR）검측성골세포（hFOB1.19）적막전철단백1（ferroportin1，Fpn1）적표체；재용불동농도적철조소（25 noml/L，50 noml/L，100 noml/L）간예배양배경，대조조가상동체적쌍증수，간예20 h，MTT법검측세포증식정황；격광공취초소묘현미경（ CLSM）검측성골세포내철리자형광염색후형광강도변화정황；Von Kossa염색검측성골세포광화정황；RT-PCR검측골개소（BGP）、I형효원（COL1）、골보호소（OPG）기인표체。결과1、성골세포존재Fpn1표체；2、철조소대성골세포증식무현저영향（ P＞0.05）；3、철조소간예후，성골세포내철리자함량증다，차여철조소간예존제량의뢰성관계（ P＜0.05）；4、철조소간예후，성골세포광화공능증강，차여철조소간예존제량의뢰성관계（ P＜0.05）；5、RT-PCR검측현시불동농도철조소간예후，각조COL1、OPG、BGP mRNA균유표체；불동농도조적COL1、OPG、BGP mRNA표체광밀도비치불동，조간밀도비치비교존재통계학의의（P＜0.05），단철조소재400 nmol/L시억제OPG적표체（P＜0.05）。결론성골세포존재철조소작용적막전철단백-1，소이철조소간예성골세포배양배경후세포내철리자발생특이성변화，동시세포광화지표현저증가、세포COL1、OPG、BGP mRNA표체함량증가；연구제시：철조소가능통과막전철단백영향성골세포철리자변화，세포철대사변화가능인기세포성골지표상조개변。
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 (25 noml/L, 50 noml/L, and 100 noml/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 Fpn 1 in osteoblasts , and the changes of iron metabolism may result in the up-regulation of osteogenesis indexes .