高铁环境对斑马鱼骨形成的影响

Effect of excess iron on bone formation in zebrafish

目的观察高铁环境对斑马鱼骨形成的影响,探讨铁过载对骨代谢影响的机制。方法以受精后第2天斑马鱼幼鱼为模型,用不同浓度枸橼酸铁铵(FAC)(0、25、50、100、200μg/mL)干预斑马鱼,受精第6天进行茜素红染色并统计骨矿化面积。在200μg/mL FAC干预6 d基础上,停止添加FAC,加入100μmol/L去铁胺(DFO),第10天行茜素红染色,统计斑马鱼椎体个数,并用实时定量PCR方法检测骨钙素(BGP)基因表达变化。结果不同浓度FAC处理后,斑马鱼骨矿化面积成剂量依赖性降低,100及200μg/mL FAC组与对照组相比差异有统计学意义(P<0.05);100μmol/L DFO加入200μg/mL FAC干预组后,斑马鱼椎体个数较未加入DFO组增加,但仍低于对照组(P<0.05);加入100μmol/L DFO组骨钙素(BGP)表达量较未加入DFO组上调,差异有统计学意义(P<0.05),但仍低于正常对照组(P<0.05)。结论高铁环境抑制斑马鱼骨形成。

Objective To explore the effect of excess iron on bone formation in zebrafish and to further investigate the effect of iron overload on bone metabolism. Methods We treated 2 days post-fertilization （2 dpf） zebrafish with different concentrations of FAC （0, 25, 50, 100, 200 μg/mL） , then Alizarin Red staining was performed at 6 dpf to analyze the bone mineralization area. Based on the FAC exposure regime, we stopped adding FAC and 100 μmol/L DFO was applied from 6dpf to 10dpf. At 10 dpf, Alizarin Red staining was performed for measuring vertebral bodies and the expression of BGP was analysed with real-time PCR. Results （ 1 ） The mineralization area was reduced in a dose-dependent-manner after FAC treatment. At the concentration of 100μg/mL and 200 μg/mL, a statistically significance was observed compared to the control group （ P 〈 0.05 ）. （2） In DFO treatment group, the number of vertebral bodies was significantly higher than that in FAC group but lower in control group （ P 〈 0.05 ）. （ 3 ） In DFO treatment group, the expression level of BGP was significantly higher than that in FAC group but lower in control group （ P 〈 0.05 ）. Conclusion Excess iron could inhibit bone formation.