南方医科大学学报

南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (01): 8-.

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活化T细胞核因子2 可抑制高迁移率蛋白1 的释放

赵擎，王黎，胡婕，刘辉   

  • 出版日期:2016-01-20 发布日期:2016-01-20

Role of nuclear factor of activated T cells-2 in high mobility protein box-1 release in
human monocytic THP-1 cells in vitro

  • Online:2016-01-20 Published:2016-01-20

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摘要： 目的探讨活化T细胞核因子2（NFAT2）对炎症因子高迁移率蛋白1（HMGB1）释放的抑制效应。方法观察脂多糖刺激可
增加HMGB1向胞外释放，在脂多糖刺激的不同时间点收集细胞及其培养上清，应用蛋白印迹法及酶联免疫法检测细胞内及培
养上清中HMGB1蛋白水平的变化；另一方面，探讨脂多糖刺激对NFAT2与HMGB1在胞浆中的结合的影响，在脂多糖刺激的
不同时间点收集THP-1细胞，提取蛋白后应用免疫共沉淀观察二者的结合情况；应用小RNA干扰技术抑制NFAT2的表达，检测
对HMGB1 合成释放的影响。结果脂多糖刺激THP-1 细胞时间越长，细胞培养上清的HMGB1 浓度增加，而细胞浆内与
NFAT2 结合的HMGB1 水平下降，细胞核内没有检测到二者的相互作用。NFAT2 的小RNA干扰质粒转染后，THP-1 细胞内
NFAT2浓度下降，同时细胞上清中HMGB1的水平上升。结论NFAT2可抑制HMGB1的合成释放。

Abstract: Obejective To investigate the role of nuclear factor of activated T cells -2 (NFAT2) in release of high mobility protein
box-1 (HMGB1) from human monocytic THP-1 cells in vitro. Methods The level of HMGB1 release from THP-1 cells in
response to lipopolysaccharide (LPS) stimulation was examined by Western blotting and enzyme-linked immunosorbent assay
(ELISA). The effect of LPS stimulation on NFAT2 and HMGB1 interaction in the cytoplasm was observed by
immunoprecipitation assay. HMGB1 production and release was detected in cells with specific small interfering RNA
(siRNA)-mediated suppression of NFAT2 expression. Results LPS stimulated HMGB1 release from THP-1 cells. As LPS
stimulation prolonged, HMGB1 concentration increased in the cell culture supernatant and decreased in the cytoplasm, and
the binding between NFAT2 and HMGB1 was not detected in the cell nuclei. NFAT2 suppression by the siRNA plasmid
resulted in increased HMGB1 level in the cell culture supernatant. Conclusion NFAT2 can inhibit HMGB1 release from THP-1
cells in vitro.