通心络对血管紧张素Ⅱ诱导的血管内皮细胞活力及组织因子的影响

Effects of Tongxinluo on cell viability and tissue factor in AngⅡ induced vascular endothelial cells

目的:研究通心络对血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(human umbilical vein endotheli-alcells,HUVECs)细胞活力(cell viability)及组织因子(tissue factor,TF)的影响和其作用机制。方法:分别用5%,10%,20%的通心络含药血浆预处理内皮细胞30min后加入AngⅡ10-6mol/L孵育HUVECs24h,观察细胞活力变化的量效关系。选择一个最佳浓度的通心络含药血浆预处理内皮细胞30min后加入AngⅡ10-6mol/L孵育HU-VECs24h观察TF,AngⅡ1型受体(AT1) mRNA水平,一氧化氮合酶(NOS)活性及一氧化氮浓度(NO),并予NOS抑制剂(L-NAME)预处理内皮细胞30min后,再加入通心络含药血浆和AngⅡ,观察孵育24h细胞活力,TF,AT1,NOS及NO变化。结果:通心络能显著提高AngⅡ诱导的血管内皮细胞活力,以10%浓度作用最明显,通心络能降低TF及AT1水平及提高NOS活性和NO浓度;L-NAME可明显抑制通心络对血管内皮的作用。结论:通心络可能通过上调NOS-NO通路提高AngⅡ诱导的内皮细胞活力及抗血栓能力。

Objective To determine the effects of Tongxinluo on cell viability and tissue factor （TF） in Ang Ⅱ induced vascular endothelial cells and to investigate its mechanism. Methods AngⅡ（ 10-6 mol/L） was added to human vascular endothelial cells （HUVECs） culture media alone or with various concentration of Tongxinluo drug containing plasma （5%, 10%, and 20% ） added 30 minutes before Ang Ⅱ. Cell viability was evaluated after 24-hour incubation with AngⅡ in a dose manner. TF, Ang Ⅱ type 1 receptor （AT1 ） mRNA, NO synthase （NOS） and NO were observed after 24-hour incubation with Ang Ⅱ. In addition, NOS inhibitor nomega-nitro-larginine （L-NAME） was added 30 minutes before Tongxinluo and Ang Ⅱ. Cell viability, TF, AT1 mRNA, the level of NOS and NO were evaluated after 24- hour incubation with Tongxinluo and Ang Ⅱ. Results Tongxinluo significantly improved Ang Ⅱ induced endothelial cell viability and the effect was the most obvious at 10%. Tongxinluo （ 10% ） decreased the TF and AT1 mRNA while increased the NOS and NO levels. L-NAME obviously inhibited the effects ofTongxinluo on cell viability, TF, AT1 mRNA, and NOS and NO levels. Conclusion Up-regulatingNOS-NO signaling may be the mechanism of Tongxinluo on cell viability and TF in Ang Ⅱ induced vacu-lar endothelial cells.