核转录因子NF-κB在血管内皮细胞组织因子表达中的作用

Effect of NF-κB on Tissue Factor Expression in Human Vascular Endothelial Cells

目的 探讨核转录因子NF-κB在肿瘤坏死因子α（TNF-α）、血管紧张素Ⅱ（AngⅡ）诱导血管内皮细胞组织因子（TF）表达中的作用。方法用发色底物法测定血管内皮细胞表面TF活性。分别采用凝胶电泳迁移率改变法(EMSA)和Western印迹法检测血管内皮细胞核转录因子NF-κB的DNA结合活性和含量。结果100 U/ml TNF-α、10-6 mol/l AngⅡ作用下，内皮细胞表面TF 活性明显增高，分别较对照增高1.65 倍和1.61 倍。TNF-α、AngⅡ作用组核抽提物与NF-κB特异DNA序列结合活性分别为正常对照组的2.07倍和3.89倍。TNF-α、AngⅡ处理后细胞核内NF-κB含量明显增加，分别为正常对照组的1.77倍和1.52倍。结论TNF-α、AngⅡ可增强内皮细胞TF活性，这可能与其促进内皮细胞核转录因子NF-κB的核转位和DNA结合活性有关，进一步证实NF-κB在内皮细胞TF诱导表达中的重要作用。

Objective To study the effect of nuclear transcription factor NF-κB on tumor necrosis factor α(TNF-α) or angiotensin Ⅱ(AngⅡ)-induced tissue factor(TF) expression in human vascular endothelial cells. Methods By chromogenic assay, we measured the TF activity of endothelial cells surface. The DNA binding activity and content of NF-κB were determined by electrophoretic mobility shift assay(EMSA) and Western blot respectively. Results TF activity of endothelial cells surface exposed to TNF-α(100 U/ml) or AngⅡ(10－6 mol/L) was significantly increased by 1.65 and 1.61 folds respectively. The binding activity of NF-κB to corresponding oligonucleotide probe in endothelial cells induced by TNF-αor AngⅡwas increased by 2.07 and 3.89 folds respectively. The content of NF-κB in endothelial cells induced by TNF-αor AngⅡwas increased by 1.77 and 1.52 folds respectively. Conclusion The increase of the content and binding activity of NF-κB may play an important role in the TNF-αor AngⅡ-induced expression of TF in endothelial cells.