黄芪、当归及川芎嗪等活血化瘀药对肝癌细胞株HepG2纤溶酶原激活物抑制剂 1表达的影响(英文)

Effect of huangqi,danggui and ligustrazine as medicines activating blood and eliminating stasis on the expression of plasminogen activator inhibitor 1 in HepG2 hepatocarcinoma cell strain

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 the main regulator of the fibrinolytic system in vivo.The increase of plasminogen activator inhibitor 1 is closely related to thrombotic disease and it is also an independent risk factor for development of thrombotic disease.OBJECTIVE:To investigate the effect of huangqi(Astragalus),danggui(Angelica) and ligustrazine as medicines activating blood and eliminating stasis on the expression of plasminogen activator inhibitor 1 in HepG2 hepatocarcinoma cell strain.DESIGN:A randomized controlled study.SETTING:First Cadre Department of General Hospital of Chinese People's Armed Police Force.MATERIALS:The experiment was completed in Academy of Military Medical Sciences of PLA from August to December 2004.HepG2 hepatocarcinoma cell strain was cultured.According to different drugs in culture medium,they were divided into six groups:control group,huangqi group,danggui group,huangqi+ danggui group,compound danshen group and ligustrazine group.METHODS:Huangqi,danggui,huangqi+ danggui,compound danshen and ligustrazine were added in HepG2 culture medium respectively.MTS assay was used to detect the effect of medicines activating blood and eliminating stasis on proliferation of HepG2 cells,plasminogen activator inhibitor 1 was assayed by specific enzyme linked immunosorbent assays(ELISA),plasminogen activator inhibitor 1 activity was measured by amidolytical assay.0.5 μ g/mL of transforming growth factor β 1 cells was added in HepG2 culture medium to stimulated production of plasminogen activator inhibitor 1.Control group was treated under the same conditions but without Chinese herbs.RESULTS:The inhibitory rates of cellular proliferation in huangqi and danggui groups were(6.51± 2.66)% and (4.42± 2.19)% ,but those in huangqi+ danggui,compound danshen and ligustrazine groups were (12.06± 4.98)% ,(16.38± 4.06)% and(32.83± 9.8)% respectively,t=2.447- 3.707,P< 0.05.Compared with the control group,huangqi,danggui,compound danshen and ligustrazine significantly inhibited plasminogen activator inhibitor 1 expression(22.68± 2.20,11.11± 1.23,19.66± 1.53,15.45± 1.27,16.90± 0.33,14.01± 0.74,t=2.447 - 3.707,P< 0.05) and activity(2.16± 0.014,2.01± 0.006,1.95 ± 0.014,1.79± 0.104,1.53± 0.045,1.48± 0.012,t=2.447- 3.707,P< 0.05) in HepG2 cells.The evident inhibitory effects were observed in the group of huangqi+ danggui,especially in compound danshen and ligustrazine.CONCLUSION:The plasminogen activator inhibitor 1 expression and activity were inhibited effectively by huangqi,danggui,compound danshen and ligustrazine.