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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 an hypoxic cell model of human umbilical vein endothelial cell line (Eahy926) and investigate the effect of ligustrazine on the fibrinolytic function of vascular endothelial cells under anoxic conditions. Methods: The cells were divided into control group, hypoxia group and ligustrazine (TMP) intervention group. The hypoxic environment of Eahy926 cells was simulated by using cobalt chloride (1.6mmol / L) to establish hypoxia model. TMP intervention group was treated with 0.08 and 0.16 g / L of TMP intervention. Hoechst method and CCK-8 method were used to detect cell viability. The expressions of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator urokinase-type plasminogen activator, u-PA) and plasminogen activator inhibitory type 1 (PAI-1) mRNA in the culture supernatants t-PA, u-PA, PAI-1 content. Results: Compared with the control group, the cell viability decreased in the hypoxia group and the TMP group (P <0.01), but compared with the hypoxia group, the cell viability increased in the TMP group (P <0.01) (P <0.01). Compared with control group, the expression of t-PA mRNA in hypoxia group was lower than that in control group (P <0.01), and the content of t-PA in cell culture supernatant (P <0.01). Compared with the hypoxia group, the expression of t-PA mRNA and the content of t-PA in the cell culture supernatant increased with the increase of TMP concentration (P <0.01) The level of t-PA mRNA was increased in low concentration of TMP intervention group (P <0.05), and the content of t-PA in cell culture supernatant was increased (P <0.01). CONCLUSION: Ligustrazine can reduce the hypoxic injury of Eahy926 cells in a concentration-dependent manner and affect the fibrinolytic function of endothelial cells by regulating the expression of t-PA mRNA and the secretion of t-PA in the supernatant.