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
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江苏省科技厅社会发展计划(BS2006007)


Effect of tetramethylpyrazine on fibrinolytic function in human umbilical vein endothelial cell (Eahy926) treated by CoCl2
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    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(1.6 mmol/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0.16 g/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 inhibitortype-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-PA mRNA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-PA mRNA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-PA mRNA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-PA mRNA表达及上清液t-PA的分泌,影响内皮细胞的纤溶功能-

    Abstract:

    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 hypoxia-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.

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吴 昊,孙 昊,王 军,黄培培,程 洁,陈旭峰,陈 彦,肖 杭,张劲松.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[J].南京医科大学学报(自然科学版),2010,(5):641-

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