川芎嗪对人脐静脉内皮细胞株(Eahy926)氯化钴诱导缺氧后纤溶功能的影响  

Effect of tetramethylpyrazine on fibrinolytic function in human umbilical vein endothelial cell(Eahy926) treated by CoCl_2

作　　者：吴昊[1] 孙昊[1] 王军[2] 黄培培[1] 程洁[2] 陈旭峰[1] 陈彦[1] 肖杭[2] 张劲松[1] 

机构地区：[1]南京医科大学第一附属医院急诊中心,江苏南京210029 [2]南京医科大学公共卫生学院神经毒理实验室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第5期641-645,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省科技厅社会发展计划(BS2006007)

摘　　要：目的:建立人脐静脉内皮细胞株(Eahy926)缺氧细胞模型,探讨川芎嗪在缺氧条件下对血管内皮细胞纤溶功能的影响。方法:细胞分为对照组、缺氧组和川芎嗪(TMP)干预组,利用氯化钴(1.6mmol/L)模拟Eahy926细胞缺氧环境,建立缺氧模型,TMP干预组分别用0.08和0.16g/L的TMP干预。Hoechst法、CCK-8法检测各组细胞活力,RT-PCR观察各组细胞组织型纤溶酶原激活物(tissue-type plasminogen activator,t-PA)、尿激酶型纤溶酶原激活物(urokinase-type plasminogen activator,u-PA)、纤溶酶原激活物抑制物-1(plasminogen activator inhibit ortype-1,PAI-1)mRNA表达,双抗体夹心ELISA法检测各组细胞培养上清液中t-PA、u-PA、PAI-1含量。结果:与对照组相比,缺氧组及TMP干预组均出现细胞活力下降(P<0.01),但与缺氧组相比,TMP干预组细胞活力上升(P<0.01),且高浓度组较低浓度组上升明显(P<0.01);RT-PCR和ELISA结果显示:与对照组相比,缺氧组t-PAmRNA表达量降低(P<0.01),细胞培养上清液t-PA含量减少(P<0.01);与缺氧组比,高低浓度TMP处理组t-PAmRNA表达量及细胞培养上清液t-PA含量均增高,具有统计学差异(P<0.01);而高浓度较低浓度TMP干预组t-PAmRNA表达量升高(P<0.05),细胞培养上清液t-PA含量增加(P<0.01)。结论:川芎嗪能够浓度依赖性减轻Eahy926细胞的缺氧损伤,并通过调节其t-PAmRNA表达及上清液t-PA的分泌,影响内皮细胞的纤溶功能。Objective：To establish hypoxia model with human umbilical vein endothelial cell line（Eahy926） and investigate the effect of tetramethylpyrazine（TMP） on Eahy926＇s fibrinolytic function with hypoxia.Methods：The cells were divided into three groups：control group,CoCl2-treated group（1.6 mmol/L） and CoCl2＋TMP-treated groups（0.08 and 0.16 g/L,respectively）.Hypoxia was mocked by treatment of CoCl2.Cell viability was evaluated by Hoechst staining and cell counting kit-8（CCK-8）;the expression of t-PA,u-PA and PAI-1 was measured by semi-quantitative RT-PCR;t-PA,u-PA and PAI-1 in the supernatant of culture medium were detected by ELISA.Results：Cell viability was decreased in CoCl2-treated and CoCl2＋TMP-treated groups compared with the control group（P 〈0.01）.Treatment of TMP improved cell viability in a dose-dependent manner（P 〈0.01）.Moreover,CoCl2 induced a decreasing mRNA expression（P 〈0.01） and secretion of t-PA（P〈 0.01）;TMP treatment elevated t-PA mRNA expression and contents of the cell supernatant（P〈 0.01） in a dose-dependent way.Conclusion：TMP can effectively protect Eahy926 cell from hy poxia-induced damage and increase viability of cells in a concentration-dependent manner;TMP might enhance the fibrinolytic function of endothelial cells by regulating the expression and secretion of t-PA.

关 键 词：川芎嗪 人脐静脉内皮细胞株(Eahy926) 氯化钴 缺氧 纤溶 

分 类 号：R364.1[医药卫生—病理学]