目的：分析丹参的主要活性成分丹参酮IIA对凝血酶诱导人脐静脉内皮细胞（HUVECs）组织因子（TF）表达的影响。方法将分离的HUVECs随机等分为空白对照组、凝血酶处理组及不同浓度丹参酮IIA处理组。其中凝血酶处理组加入凝血酶（终浓度5U/ml），不同浓度丹参酮IIA处理组加入凝血酶（终浓度5U/ml）和不同浓度的丹参酮IIA（终浓度分别为0.25μg/ml、0.5μg/ml、0.75μg/ml和1.0μg/ml），空白对照组则加入等量的无血清培养基。各组HUVECs均在37℃培养6h后收集细胞。结果各组HUVECs孵育6h后，凝血酶处理组HUVECs中TF mRNA的转录明显强于空白对照组（P<0.001），不同浓度的丹参酮IIA可不同程度抑制TF mRNA的转录，与凝血酶处理组比较差异均有统计学意义（P<0.001），且呈剂量依赖关系。HUVECs与凝血酶孵育后，所表达的TF促凝活性（TF：C）和抗原含量（TF：Ag）均明显增加，明显高于空白对照组（P<0.001）。不同浓度的丹参酮IIA均可不同程度地抑制凝血酶所诱导的TF：C和TF：Ag表达（P<0.001），且呈剂量依赖关系。结论丹参酮IIA可抑制凝血酶诱导的人脐静脉内皮细胞TF基因的转录和表达，这可能是丹参酮IIA发挥抗血栓形成作用的重要机制。%Objective The effect of tanshinone IIA(Tan IIA),the main active component from traditional Chinese herb-Radix Salviae Miltiorrhizae,on thrombin induced tissue factor(TF)expression in human umbilical vein endothelial cells (HUVECs)was investigated. Methods Cultured HUVECs were randomly divided into control group,thrombin-treated group and Tan IIA-treated groups with different concentrations of Tan II A. In thrombin-treated group,HUVECs were incubated with thrombin (final concentration of 5U/ml)for 6 hours; while in Tan IIA-treated groups,HUVECs were incubated with thrombin(final concentration of 5U/ml)plus Tan IIA with the final concentration of 0.25μg/ml,0.5μg/ml,0.75μg/ml and 1μg/ml respectively for 6 hours;in control group,no thrombin and Tan II A was added. After incubation for 6 hours at 37℃,HUVECs in all groups were harvested. Results After incubation for 6 hours at 37℃,there was only trace of TF mRNA existed in HUVECs from control group, while TF mRNA from HUVECs of thrombin-treated group was augmented obviously than that in control group(P<0.001),and Tan IIA with different concentrations inhibited the transcription of TF mRNA in different degree with a dose-dependent manner(P<0.001, compared with thrombin-treated group). After incubation with thrombin,both TF procoagulant activity(TF:C)and TF antigen (TF:Ag)expressed in HUVECs increased markedly(P<0.001,compared with control group),while they were inhibited by Tan IIA with different concentrations in a dose-dependent manner in Tan IIA-treated groups(P<0.001,compared with thrombin-treated group). Conclusion Tanshinone IIA inhibits the transcription and expression of TF gene in human umbilical vein endothelial cells induced by thrombin,this may be one of the mechanisms of Tanshinone IIA antagonizing thrombosis.
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