【摘要】 目的观察肿瘤坏死因子(TNFα)对人脐静脉血管内皮细胞株ECV304组织因子(TF)表达的影响及植物来源单体化合物川芎嗪的干预作用。方法内皮细胞培养采用RPMI-1640完全培养基;采用一期凝固法测总的细胞促凝活性(PCA);细胞TF抗原测定采用ELISA法;TFmRNA采用RT-PCR的方法检测。结果川芎嗪预处理后,可明显抑制TNFα(1000u/mL)作用不同时间以及不同浓度的TNFα(0～2000u/mL)作用6h后ECV304细胞TF活性的表达;川芎嗪在1～1000μg/mL范围内以剂量依赖方式抑制TN-Fα(1000u/mL)的刺激作用(P<0.01),在1000μg/mL时,川芎嗪的抑制作用最强;以其预处理ECV304,可使TNFα刺激TF的活性下降70%;川芎嗪也可显著降低受刺激的ECV304TF抗原及TFmRNA表达。结论TNFα可诱导血管内皮细胞TF的表达,川芎嗪具有在mRNA水平抑制TNFα刺激血管内皮细胞TF表达的作用。更多还原

【Abstract】 Objective Tetramethylpyrazine (TMP) is a Chinese plant-derived compound used as antithrombotic drug. The purpose of this study was to observe the effect of TMP on the expression of tissue factor (TF) induced by tumor necrosis factor (TNF)α in human umbilical vein endothelium derived cell line ECV304. Methods ECV304 was cultured in RPMI-1640. TF activity was determined with one-stage clotting assay measuring total cellular pro-coagulant activity. TF antigen was assayed by ELISA. TF mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Results The preincubation of TMP inhibited TF activity induced by different hours or different concentrations of TNFα. Increasing concentrations of TMP (1~1000 μg/mL) reduced the TF activity,TF antigen induced in ECV304 by TNFα (1000 u/mL) (P <0.01),in a dose-dependent fashion. TF activity induced by TNFα decreased to approximately 30% after treating with 1000 μg/mL of TMP. TMP also reduced the expression of TF mRNA induced by TNFα. Conclusion The present data suggests that TNFα induced the TF expression of vascular endothelial cells in a dose and time-dependent fashion,and TMP could inhibit this effect at mRNA level.更多还原