活性氧在铁过载影响成骨细胞生物活性中的作用  

Function of reactive species in effect of iron overload on the biological activity of osteoblasts

作　　者：何银锋[1] 高超[1] 赵国阳[1] 张林林[1] 张增利[2] 林华[3] 徐又佳[1] 

机构地区：[1]苏州大学附属第二医院骨科,苏州215004 [2]苏州大学公共卫生学院维生素D研究室,苏州215007 [3]南京大学附属鼓楼医院骨科,南京10008

出　　处：《中国骨质疏松杂志》2013年第6期541-546,共6页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金资助项目(81273090);江苏省自然科学基金资助项目(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(SZS201208);苏州市应用基础研究计划之医疗卫生部分资助项目(SYS201002)

摘　　要：目的观察铁过载对人成骨细胞(hFOB1.19)生物活性的影响,同时观察活性氧在这一实验变化过程中的作用。方法体外培养成骨细胞,一组运用200μmol/L枸橼酸铁铵(FAC)干预,一组运用2.5 mmol/L抗氧化剂N-乙酰半胱氨酸(NAC)干预,一组NAC预处理1 h后运用相同浓度FAC干预,一组为正常对照;细胞培养48 h后,流式细胞仪检测各组细胞内活性氧(ROS)的水平;CCK-8法检测各组细胞活力;RT-PCR法检测各组细胞OPG、BGP和COL1 mRNA表达的变化;碱性磷酸酶活性试剂盒检测各组细胞碱性磷酸酶活性。结果不同干预组成骨细胞内活性氧含量差异显著不同(P<0.05),FAC组显著高于对照组,FAC+NAC组低于FAC组、高于NAC组;各组间活性氧含量的变化与成骨细胞活力、OPG、BGP和COL1 mRNA表达光密度比值、碱性磷酸酶活性呈负相关性,组间比较有统计学差异(P<0.05)。结论铁过载降低成骨细胞生物活性可能与铁过载增加成骨细胞内活性氧水平有关。Objective To observe the effect of iron overload on the biological activity of human osteoblasts （hFOB1.19） in vitro, and to observe the function of reactive species in this progress. Methods Osteoblasts were cultured in vitro and divided into 4 groups. One group was treated with 200 μmol/L ferric ammonium citrate （FAC） ; one group was treated with 2.5 mmol/L antioxidant N-aeetyl cysteine （NAC） ; one group was pretreated with NAC for lh and then treated with the same concentration of FAC intervention; and the last group was normal control group. After 48 - hour culturing, the levels of reactive oxygen species （ROS） in each group were detected using flow cytometry. Cell viability was detected using CCK- 8 assay. The expression of OPG, BGP, and COL1 mRNA was detected using RT-PCR. Alkaline phosphatase （ALP） activity was detected using ALP viability kit. Results The levels of reactive oxygen species among different groups were significantly different （P 〈 0. 05）. The level in FAC group was higher than that in control group, and the level in FAC ＋ NAC group was significantly lower than that in FAC group, but higher than that in NAC group. The content of active oxygen in each group was negatively correlated with osteoblast activity, the optical density ratio of OPG, BGP, and COLlmRNA expression, and alkaline phosphatase activity （ P 〈0. 05 ）. Conclusion The effect of iron overload on reducing the biological activity of osteoblasts may be associated with increased reactive oxygen species caused by iron overload.

关 键 词：人成骨细胞 铁过载 活性氧 细胞活性 

分 类 号：R363[医药卫生—病理学]