金属离子刺激单核-巨噬细胞核因子κB受体活化子的研究  

Effect of Metal Ions on Monocyte-Macrophage Cells

作　　者：陈锐[1] 戴闽[1] 帅浪[1] 张斌[1] 付文锋[1] 熊皞[1] 

机构地区：[1]南昌大学第一附属医院骨二科,330006

出　　处：《天津医药》2010年第10期874-877,共4页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目(项目编号:30760255)

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-巨噬细胞表达核因子κB受体活化子(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,不同时间用噻唑蓝(MTT)比色法检测其细胞活性。将细胞分为5组,A组为单纯单核-巨噬细胞,B组为单核-巨噬细胞+500mg/LCr3+,C组为单核-巨噬细胞+500mg/LCr3++20μmol/LSP600125,D组为单核-巨噬细胞+10mg/LCo2+,E组为单核-巨噬细胞+10mg/LCo2++20μmol/LSP600125。用半定量逆转录-聚合酶链反应(RT-PCR)在24h、48h检测各组细胞的RANK mRNA的表达量。结果:与A组相比,B组和D组单核-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可抑制Cr3+、Co2+刺激的单核-巨噬细胞RANK mRNA表达(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。Objective：To investigate the cytotoxicity and macrophage expression of receptor activator of nuclear factor κB（RANK） induced by chromium（Cr3＋） and cobalt（Co2＋） ions,and the mechanism of osteolysis after artificial joint replacement.Methods：The monocyte-macrophage cells were cultured in vitro and 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＋ 500 mg/L Cr3＋）,Group C（monocyte /macrophages＋500 mg/L Cr3＋＋ 20 μmol/L SP600125）,Group D（monocyte /macrophages＋10 mg/L Co2＋） and Group E（monocyte /macrophages ＋10 mg/L Co2＋ ＋20 μmol/L SP600125）.The semi-quantitative reverse transcription PCR method was used to detect the level of RANK mRNA after 24 h and 48 h.Results：MTT test demonstrated that the cell viability of monocyte /macrophages was obviously decreased by Co2＋ and Cr3＋ ions in B and D groups than that of A group.The mRNA expression of RANK was increased in cells after exposed to Co2＋ and Cr3＋ ions for 24 h and 48 h（P 0.05）.This mRNA expression of RANK stimulated by Co2＋ and Cr3＋ was inhibited by SP600125（JNK Inhibitor） in monocytemacrophage cells（P 0.05）.Conclusion：Co2＋ and 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.

关 键 词：铬 钴 单核巨噬细胞系统 NF-ΚB 受体 胞质和核 原癌基因蛋白质c-jun 人工关节 逆转录聚合酶链反应 

分 类 号：R382.31[医药卫生—医学寄生虫学]