氟伐他汀对TNF-α诱导的组织因子活性与表达的影响

The effects of fluvastatin on the expression and activity of TNF-α-induced tissue factor

目的研究不同浓度氟伐他汀(fluvastatin,Flu)对由肿瘤坏死因子-α(TNF-α)诱导的人脐静脉内皮细胞(HUVECs)产生组织因子(tissue factor,TF)含量、活性及TF mRNA表达、TF蛋白水平的影响,探讨他汀类药物在抗血栓形成方面的作用。方法将同一簇HUVECs进行培养传代,分设空白组、TNF-α组、Flu组及干预组(TNF-α+Flu组),其中TNF-α+Flu组中分别以不同浓度Flu干预,采用ELISA方法测定TF含量,用发色底物法测定TF活性,用RT-PCR方法测定TF mRNA表达水平,用Western-blot方法测定TF蛋白表达。结果 TNF-α组TF含量、TF活性、TF RNA表达和TF蛋白水平均较空白组增高(P<0.05);与TNF-α组比较,不同浓度Flu干预组TF含量与活性以及TF mRNA表达和TF蛋白水平均明显降低,差异有显著性(P<0.05);Flu的抑制作用在1μmol/L以内时呈浓度依赖性,可显著抑制由TNF-α诱导的TF水平及活性(P<0.05),当Flu浓度>1μmol/L,作用差异无显著性(P>0.05)。结论 Flu可有效抑制由TNF-α诱导的人脐静脉内皮细胞TF含量、活性,TF mRNA、蛋白表达水平,提示Flu具有潜在的降脂之外的抗动脉血栓作用。

Objective To investigate the effects of various concentrations of Fluvastatin （Flu） on the content, activity, and mRNA and protein expression levels of TNF-α -induced tissue factor （TF） in human umbilical vein endothelial cells （ HUVECs）, thus to evaluate the effect of statins on anti - thrombosis formation. Methods The single strain HUVECs were cultured and passaged into control group, TNF-α group, Flu group, and TNF -α ＋ Flu group. In TNF-α ＋ Flu group, cells were subdivided into subgroups according to different concentrations of Flu added. TF content, activity, mRNA expression and protein expression were assessed with ELISA, chromatogenous substrate assay, RT - PCR and Western - Blot, respectively. Results Significant increase of TF content, activity, mRNA expression, and protein level were revealed in TNF -α group, when comparing with those in control group （ P 〈0. 05 ）. However, all these were signif- icantly inhibited by Flu intervention （ P 〈 0. 05 ） ;This effect was following a significantly concentration - dependent manner when Flu was lower than 1μmol/L （ P 〈 0.05 ）. Conclusion Flu inhibits the TNF-α - induced TF content, activity, and mRNA and protein expression levels effectively in HUVECS, suggesting a potential novel external anti - thrombosis application of Flu.