[核因子-κB诱饵对培养的人脐静脉内皮细胞组织因子基因表达及凝血因子VII激活的抑制作用研究]

[Study of the inhibitory effect of NF-kappaB decoy on tissue factor gene expression and FVII activation in cultured human umbilical vein endothelial cells].

作者信息

Wang Lin-lin, Wei Wen-ning, Hu Yu, Song Shan-jun

机构信息

Institute of Hematology, Tongji Medical College, Huazhong Science & Technology University, Wuhan 430022, China.

出版信息

Zhonghua Xue Ye Xue Za Zhi. 2003 Mar;24(3):149-51.

PMID:12697128

Abstract

OBJECTIVE

To investigate the inhibitory effect of NF-kappaB decoy on tissue factor (TF) expression and FVII activation in cultured human umbilical vein endothelial cells (HUVEC), and to explore new methods for prevention and treatment of coronary heart disease.

METHODS

NF-kappaB decoy transfection efficiency was detected by flow cytometry, NF-kappaB decoy's mechanism was analyzed by electrophoretic mobility shift assay (EMSA), TF mRNA was detected by RT-PCR, TF antigen expression on the surface of HUVEC by flow cytometry, FVIIa level in plasma incubated with HUVEC stimulated by TNF-alpha by rsTF one stage clotting method.

RESULTS

NF-kappaB decoy could be successfully transfected into HUVEC. It could compete with the endogenous kappaB cis sequence element in the regulatory regions of TF promoter to bind transcriptional factor NF-kappaB. It could also significantly inhibit the TF mRNA, TF antigen expression on the cell surface and TF function leading to activation of FVII.

CONCLUSION

NF-kappaB decoy could inhibit TF gene expression and FVII activation in cultured HUVEC and might be a potential new strategy for prevention and treatment of coronary heart disease.

摘要

目的

研究核因子κB（NF-κB）诱饵对培养的人脐静脉内皮细胞（HUVEC）中组织因子（TF）表达及凝血因子VII（FVII）激活的抑制作用，探索冠心病防治的新方法。

方法

采用流式细胞术检测NF-κB诱饵的转染效率，采用电泳迁移率变动分析（EMSA）分析NF-κB诱饵的作用机制，采用逆转录-聚合酶链反应（RT-PCR）检测TF mRNA，采用流式细胞术检测HUVEC表面TF抗原表达，采用重组可溶性组织因子（rsTF）一步法凝血检测经肿瘤坏死因子-α（TNF-α）刺激的HUVEC孵育血浆中的FVIIa水平。