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目的观察中药黄芪、当归对血管内皮细胞纤溶酶原激活物抑制剂-1(plasminogen activator inhibitor-1,PAI-1)表达及活性的影响,探讨黄芪、当归抗血栓作用的分子机制。方法使用成品的黄芪和当归注射液,各设两个浓度(6mg/ml和3mg/ml),分别加入细胞培养板,实验组各孔均加入0.5ng/ml TGF-β1作为诱导剂以增加PAI-1Ag表达。同时设立TGF-β1组(细胞液中只加入0.5ng/ml TGF-β1)以及空白对照组(细胞液中不加入任何成分),各组在37℃,5%CO2孵箱中培养24h。应用RT-PCR方法检测黄芪、当归对血管内皮细胞PAI-1mRNA表达的影响,ELISA法检测黄芪、当归对PAI-1表达的影响,发色底物显色法检测黄芪、当归对PAI-1活性的影响。结果黄芪、当归可不同程度地降低血管内皮细胞PAI-1mRNA表达、抗原水平与活性,以黄芪、当归联合应用作用更为明显。黄芪、当归单独及联合应用对PAI-1mRNA表达的抑制率分别为17%、24.6%和36.6%。结论黄芪、当归可能通过抑制血管内皮细胞PAI-1表达和活性而发挥其抗血栓形成的作用。
Objective To observe the effect of Astragalus membranaceus and Angelica sinensis on expression and activity of plasminogen activator inhibitor-1 (PAI-1) in vascular endothelial cells and explore the molecular mechanism of anti-thrombus effect of Astragalus and Angelica sinensis. Methods The finished products of Astragalus membranaceus and Angelica sinensis were injected into the cell culture plate at two concentrations (6mg / ml and 3mg / ml respectively). Each group in the experimental group was given 0.5ng / ml TGF-β1 as an inducer to increase PAI -1Ag expression. At the same time, TGF-β1 group (0.5ng / ml TGF-β1) and blank control group (without adding any components) were set up in each group, and each group was cultured at 37 ℃ in 5% CO2 incubator for 24h. The effects of Astragalus membranaceus and Angelica sinensis on the expression of PAI-1 mRNA in vascular endothelial cells were detected by RT-PCR. The effect of Astragalus membranaceus and Angelica sinensis on the expression of PAI-1 was detected by ELISA. The activity of PAI-1 Impact. Results Astragalus membranaceus and Angelica sinensis could reduce PAI-1mRNA expression, antigen level and activity in vascular endothelial cells to a certain extent, especially in the combination of Astragalus membranaceus and Angelica sinensis. Astragalus membranaceus and Angelica sinensis could inhibit PAI-1 mRNA expression by 17%, 24.6% and 36.6%, respectively. Conclusion Astragalus and Angelica can exert their antithrombotic effect by inhibiting the expression and activity of PAI-1 in vascular endothelial cells.