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金属离子刺激单核—巨噬细胞核因子kB受体活化子的研究

陈锐1,戴闽2,帅浪3,张斌3,付文锋3,熊皞3   

  1. 1. 南昌大学第一附属医院骨二科
    2. 南昌大学第一附属医院
    3.
  • 收稿日期:2009-11-26 修回日期:2010-05-13 出版日期:2010-10-15 发布日期:2010-10-15
  • 通讯作者: 陈锐

The related mechanism of artificial joint loosening caused by monocyte-macrophage cells challenged with metal ions

  • Received:2009-11-26 Revised:2010-05-13 Published:2010-10-15 Online:2010-10-15

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摘要： 摘要：[目的] 探讨Cr3+ 、Co2+对体外培养的小鼠单核/巨噬细胞(RAW264.7) 的细胞毒性以及刺激单核/巨噬细胞表达RANK在人工关节术后引起骨溶解的分子生物学机制。[方法] 在体外培养单核/巨噬细胞(RAW264.7)。用Co2+、Cr3+分别干预单核/巨噬细胞，不同时间用MTT方法检测其细胞活性。将细胞分为5组：A:单纯单核/巨噬细胞; B:单核/巨噬细胞+500 ppm 铬离子;C:单核/巨噬细胞+500ppm铬离子+ 20μΜ SP600125;D:单核/巨噬细胞+10ppm 钴离子;E:单核/巨噬细胞+10ppm 钴离子+ 20μΜ SP600125; 用半定量逆转录-聚合酶链反应(RT-PCR)方法在24h、48h检测各组细胞的RANK mRNA 的表达量。[结果] MTT显示与对照组相比，钴和铬离子都能使单核/巨噬细胞的细胞活力明显下降。在24h、48h时，Co2+ 和Cr3+均能刺激单核/巨噬细胞使其细胞RANK mRNA表达量增强（P<0.05）且SP600125（JNK抑制剂）可抑制钴和铬离子刺激单核/巨噬细胞表达RANK mRNA（P<0.05）。[结论]金属离子对单核/巨噬细胞有细胞毒性,且能够诱导单核/巨噬细胞RANK mRNA 的表达，此外JNK通路参与金属离子刺激单核/巨噬细胞表面RANK的表达。

关键词: 人工关节, 铬离子, 钴离子, 单核巨噬细胞, RANK

Abstract: Abstract: Objective Research the cytotoxicity of chromium (Cr3+) and cobalt(Co2+) ions and the mechanism of osteolysis after artificial joint replacement by RANK express in rats monocyte/macrophages(RAW264.7) stimulated with Co2+ and Cr3+. Methods Culture monocyte-macrophage cells in vitro. Monocyte-macrophage cells exposed to Co2+ and Cr3+ ions, The cell viability was assured by MTT test. The cells were divided into 5 groups, Group A: monocyte /macrophages; Group B: monocyte /macrophages + 500ppm Cr3+ ;Group C: monocyte /macrophages +500ppm Cr3++ 20μM SP600125; Group D: monocyte /macrophages + 10ppm Co2+; Group E: monocyte /macrophages + 10ppm Co2+ + 20μM SP600125 ; Then,Semi-quantitative reverse transcription PCR method was used to detect the level of RANK mRNA after 24h and 48h. Results Compared to the control, MTT test demonstrated that Co2+and Cr3+ ions can both obviously decrease the cell viability of monocyte /macrophages. Cells exposed to Co2+ and Cr3+ ions，compared to the control（Group A），mRNA expression of RANK was both incresaed at 24h and 48h（P<0.05）,and SP600125 (JNK Inhibitor) could inhibite cells to express of RANK mRNA in cells stimulated with Co2+ and Cr3+（P<0.05）. Conclusion Co2+、Cr3+ ions have a cytotoxic effect on monocyte/macrophages and can stimulate the expression of RANK in monocyte /macrophages.In addition,metal ions could stimulate monocyte /macrophages to express RANK through JNK pathway.

Key words: artificial joint, cobalt ion, column ion, monocyte macrophages, RANK