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背景:纤溶酶原激活物抑制剂1是体内纤溶状态的重要调节者,其升高与血栓形成密切相关,是形成血栓性疾病的独立危险因素。目的:观察活血化瘀药黄芪、当归、复方丹参、川芎嗪对肝癌细胞株HepG2细胞增殖和纤溶酶原激活物抑制剂1表达及活性的影响。设计:完全随机设计,对照实验。单位:武装警察部队总医院干一科。材料:实验于2004-08/2004-12在军事医学科学院完成。培养肝癌细胞株HepG2细胞,根据培养液中加入的干预药物不同而分为6组:对照组、黄芪组、当归组、黄芪+当归组、复方丹参组、川芎嗪组。方法:将中药黄芪、当归、黄芪+当归、复方丹参、川芎嗪分别加入对体外培养的肝癌细胞株HepG2细胞,应用四氮唑蓝比色方法观察对肝癌细胞株HepG2细胞增殖的影响;应用采用酶联免疫吸附法测定观察活血化瘀药对细胞内纤溶酶原激活物抑制剂1表达的影响,发色底物显色法检测法检测对纤溶酶原激活物抑制剂1活性的影响。实验组各孔均加入0.5μg/L转化生长因子β1作为诱导剂以增加纤溶酶原激活物抑制剂1抗原表达。对照组细胞液中只加入0.5μg/L转化生长因子β1。结果:①加药24h后,黄芪、当归组肝癌细胞株HepG2细胞增殖抑制率分别为(6.51±2.66)%,(4.42±2.19)%;黄芪+当归组、复方丹参组、川芎嗪组肝癌细胞株HepG2增殖抑制率分别为(12.06±4.98)%,(16.38±4.06)%,(32.83±9.8)%,t=2.447～3.707,P<0.05。②黄芪、当归单独及联合应用、复方丹参、川芎嗪组HepG2细胞纤溶酶原激活物抑制剂1抗原水平明显低于对照组(17.11±1.23,19.66±1.53,15.45±1.27,16.90±0.33,14.01±0.74,22.68±2.20,t=2.447～3.707,P<0.05)。③黄芪、当归单独及联合应用、复方丹参、川芎嗪组肝癌细胞株HepG纤溶酶原激活物抑制剂1活性明显低于对照组(2.01±0.006,1.95±0.014,1.79±0.104,1.53±0.045,1.48±0.012,2.16±0.014,t=2.447～3.707,P<0.05)。以黄芪当归联合应用和复方丹参、川芎嗪作用更为明显。结论:黄芪、当归、复方丹参、川芎嗪在细胞水平能够有效抑制纤溶酶原激活物抑制物1表达和活性。
BACKGROUND: Plasminogen activator inhibitor-1 is an important regulator of fibrinolytic status in the body and its elevation is closely related to thrombosis and is an independent risk factor for the formation of thrombotic diseases. Objective: To observe the effects of Huangqi, Angelica, Salvia miltiorrhiza and ligustrazine on the proliferation and activity of plasminogen activator inhibitor 1 in HepG2 cells. Design: completely random design, control experiment. Unit: Armed Police General Hospital, a dry branch. Materials: The experiment was performed at the Academy of Military Medical Sciences from August 2004 to December 2004. The HepG2 cells were cultured and divided into 6 groups according to different intervention drugs in the culture medium: control group, Astragalus membranaceus group, Angelica group, Astragalus + Angelica group, Fufang Danshen group and ligustrazine group. Methods: Astragalus membranaceus, Angelica sinensis, Radix Astragali, Angelica sinensis, Radix Salviae Miltiorrhizae and ligustrazine were respectively added to HepG2 cells cultured in vitro. The effect of HepG2 cells proliferation was observed by tetrazolium blue colorimetric method. Enzyme-linked immunosorbent assay was used to observe the effect of Huoxue Huayu decoction on the expression of plasminogen activator inhibitor-1 and the effect of chromogenic substrate assay on the activity of plasminogen activator inhibitor-1 . 0.5μg / L transforming growth factor β1 was added into each well of experimental group as an inducer to increase plasminogen activator inhibitor-1 antigen expression. In the control group, only 0.5μg / L transforming growth factor β1 was added into the cytosol. Results: ①The inhibitory rates of proliferation of HepG2 cells were (6.51 ± 2.66)% and (4.42 ± 2.19)%, respectively, in astragalus membranaceus and Angelica sinensis The inhibition rate of strain HepG2 was (12.06 ± 4.98)%, (16.38 ± 4.06)%, (32.83 ± 9.8)%, t = 2.447 ~ 3.707, P <0.05. Astragalus membranaceus and Angelica sinensis could be used alone and in combination. The level of plasminogen activator inhibitor-1 in HepG2 cells of compound Danshen and ligustrazine groups was significantly lower than that of the control group (17.11 ± 1.23, 19.66 ± 1.53, 15.45 ± 1.27, 16.90 ± 0.33, 14.01 ± 0.74,22.68 ± 2.20, t = 2.447 ~ 3.707, P <0.05). ③ Astragalus membranaceus and Angelica sinensis were used alone and in combination. The activity of HepG plasminogen activator inhibitor 1 in compound Danshen and ligustrazine groups was significantly lower than that in the control group (2.01 ± 0.006, 1.95 ± 0.014, 1.79 ± 0.104, 1.53 ± 0.045 , 1.48 ± 0.012, 2.16 ± 0.014, t = 2.447 ~ 3.707, P <0.05). Astragalus angelica combined application and compound Salvia, ligustrazine effect is more obvious. Conclusion: Astragalus membranaceus, Angelica sinensis, Radix Salviae Miltiorrhizae and ligustrazine can effectively inhibit plasminogen activator inhibitor-1 expression and activity at the cellular level.