南方医科大学学报

南方医科大学学报 ›› 2006, Vol. 26 ›› Issue (01): 102-104.

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寡聚脱氧核苷酸抑制巨噬细胞NF-κB活性的实验研究

崔忠林;周杰;   

  1. 南方医科大学南方医院肝胆外科； 南方医科大学南方医院肝胆外科 广东 广州 510515； 广东 广州 510515；
  • 出版日期:2006-01-20 发布日期:2006-01-20
  • 基金资助:
    广东省自然科学基金(960327)

Inhibition of nuclear translocation and binding activity of nuclear factor-κB by oligodeoxynu-cleotides in THP-1 cells

CUI Zhong-lin, ZHOU Jie Department of Hepatobiliary Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China   

  1. 南方医科大学南方医院肝胆外科； 南方医科大学南方医院肝胆外科 广东 广州 510515； 广东 广州 510515；
  • Online:2006-01-20 Published:2006-01-20

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摘要： 目的观察寡聚脱氧核苷酸(ODN)对THP-1细胞NF-κB核转位及细胞核内结合活性的抑制作用。方法用脂质体将寡聚脱氧核苷酸转入细胞后,以细菌脂多糖(LPS)刺激30min,分别通过免疫细胞化学、电泳迁移率分析(EMSA)和 RT-PCR检测细胞NF-κB的核移位和细胞核内NF-κB结合活性。结果免疫细胞化学显示转入ODN后以LPS刺激, NF-κB表达仍位于细胞浆内,EMSA显示ODN可明显抑制细胞核内结合活性,RT-PCR显示ODN能够明显抑制 NF-κB相关炎性因子TNF-α mRNA的合成。结论ODN能有效抑制THP-1细胞NF-κB的核移位和细胞核内NF-κB 结合活性,从而抑制相关炎性基因的启动和转录。 更多还原

Abstract: Objective To observe the inhibitory effect of oligodeoxynucleotides (ODN) on nuclear translocation and nuclear binding activity of nuclear factor (NF)-KB in THP-1 cells. Methods Oligodeoxynucleotides were tansfected via liposome into THP-1 cells followed by stimulation of the cells with lipopolysaccharide (LPS). Immunocytochemistry, electrophoretic mobility shift assay (EMSA) and reverse transcription (RT)-PCR were performed to detect the nuclear translocation and nuclear binding activity of NF-κB. Results Immunocytochemical results showed that after LPS stimulation of the ODN-transfected cells, NF-κB expression was still localized in cytoplasma. EMSA demonstrated inhibited nuclear binding activity of NF-κB in the ODN-transfected cells, and ODN inhibited the mRNA expression of tumor necrosis factor (TNF)-α, a NF-κB-associated inflammatory factor, as shown by RT-PCR. Conclusion ODN can inhibit the nuclear translocation and binding activity of NF-κB in THP-1 cells, whereby the transcription and expression of the related inflammation factor genes is suppressed, which shed light on a new solution for clinical treatment of acute pancreatitis.

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