TNF-α刺激的人脐静脉内皮细胞组织因子表达及其分子机制

Tissue Factor Expression in Human Umbilical Vien Endothelial Cells Stimulated by TNF-α and Its Molecular Mechanism

为探讨人脐静脉内皮细胞 (HUVEC)在肿瘤坏死因子 (TNF α)刺激下组织因子 (TF)的表达及其分子机制 ,用流式细胞术检测HUVEC膜表面TF蛋白的表达 ,用RT PCR方法检测TFmRNA的表达和用Western印迹方法检测HUVEC核蛋白中核因子κB(NF κB)。结果发现 ,TNF α能刺激HUVEC细胞膜表面TF表达增加 ,其增加与TFmRNA合成增加具有高度一致性 ,且TF表达的增高滞后于mRNA升高数小时之后 ;TF表达增加的同时伴有核蛋白 (κB)显著的活化。结论 :TNF α刺激HUVEC后 ,迅速活化NF κB ,启动TF的转录 ,TFmRNA表达增加 ,TF蛋白合成增加 ,导致TF在HUVEC膜表面表达增加 。

The objective of this study was to explore tissue factor (TF) expression induced by TNF α in cultured human umbilical vien endothelial cells (HUVEC) and its molecular mechanism. TF expression on the surface of HUVEC, TF mRNA and nuclear factor κB (NF κB) in HUVEC were detected by flow cytometry, RT PCR and Western blot respectively. The results showed that TNF α could enhance TF expression on the surface of HUVEC, the TF expression increase was highly consisitent with the increased synthesis of TF mRNA, and the increase of TF expression was lately appeared for several hours. It was also found activation of NF κB at the time TF mRNA increase. In conclusion, NF κB could be activated promptly after HUVEC incubated with TNF α, then it was bound to TF promotor to start the TF transcription, TF mRNA expression was upregulated, that leaded to the increase of TF expression on the HUVEC surface and activated the coagulation cascade.