目的应用人脐静脉内皮细胞(HUVECs)缺氧模型,观察川芎嗪对其抗凝和纤溶功能的影响。方法制备正常人脐静脉原代EC,待EC培养达融合状态后,将其放入含95%N2、5%CO₂混合气体的缺氧环境,制备人脐静脉原代EC缺氧损伤模型。并设正常组,模型组,川芎嗪组,观察各组细胞培养上清液中漂浮细胞数、抗凝、纤溶指标等变化。结果与对照组相比,模型组培养液中的漂浮细胞数显著增高,前列环素(6-keto-PGF1α)水平显著下降,组织型纤溶酶原激活物抑制物(PAI)活性显著增高,组织型纤溶酶原激活物(t-PA)活性显著降低,内皮素(ET)水平显著增高,NO水平显著降低;川芎嗪具有显著提高6-keto-PGF1α水平,调节PAI、t-PA活性和ET、NO水平的作用。结论本实验制备的人脐静脉原代EC缺氧损伤模型存在着EC受损,分泌功能下降,血管舒缩功能失调;川芎嗪具有保护在缺氧环境下EC受损,调节EC分泌功能和血管舒缩功能。

Objective To set the hypoxic model in human umbilical vein endothelium cells (HUVECs) and observe the effect of tetramethylpyrazine in its functions of anticoagulation and fibrinolysis. Methods Preparing the normal primary HUVECs, when the cultured endothelial cells (ECs) were inosculated they were put into the anoxic condition with 95% N₂ and 5% CO₂ to make hypoxic model in primary HUVECs. Normal, model, and tetramethylpyrazine groups were divided and used to observe the difference of floating cells, anticoagulation and fibrinolysis indexes in supernatant of each group. Results Compared with the control group, the number of floating cells in supernatant was increased obviously, the level of 6-keto-PGF1α was declined significantly, the activity of plasminogen activator inhibitor (PAI) and the level of endothelin (ET) were enhanced obviously, the activity of tissue plasminogen activator (t-PA) and the level of NO were declined significantly in model group; the level of 6-keto-PGF1α was raised significantly, the activity of PAI and t-PA, and the levels of ET and NO could be regulated in the tetramethylpyrazine group. Conclusion In the hypoxic model of primary HUVECs made in this experiment, ECs are injured, secreting function is declined, vasomotion is out of control. Tetramethylpyrazine could protect hypoxic ECs from being injured, regulate the secreting function of ECs and the vasomotion. The model could provide a method for basic study and medicine selection of anticoagulation and fibrinolysis.

国家自然科学基金资助项目(30371789)；浙江省自然科学基金资助项目(Z204424)