蕲蛇酶对血管内皮细胞纤维蛋白溶解功能的影响

目的　研究蕲蛇酶对体外培养人脐静脉内皮细胞纤溶活性的影响 ,探讨其抗栓作用机制。方法　采用人脐带来源的脐静脉内皮细胞进行原代及传代培养 ,以光镜、电镜、第Ⅷ因子相关抗原免疫组化等方法鉴定内皮细胞 ,采用发色底物法 (S2 2 51 )测定内皮细胞培养液中组织型纤溶酶原激活剂(t PA)及组织型纤溶酶原激活剂抑制物 (PAI)的活性 ,ELISA法测定培养液中纤维蛋白降解产物的含量。结果　所培养的细胞具有特征性Weibel palade小体 ,第Ⅷ因子特异性抗原阳性。蕲蛇酶使细胞培养液中t PA活性升高 ,t PA/PAI比值升高 ,纤维蛋白降解产物明显增加。结论　蕲蛇酶具有促进血管内皮细胞释放t PA ,提高其纤溶活性的作用。

Effect of acutobin on the activity of fibrinolysis in cultured human umbilical vein endothelial cells

AIM This study is to observe the effects of acutobin on the activity of tissue type plasminogen activitor(t PA) and tissue plasminogen activitor inhibitor(PAI) in the cultured human umbilical vein endothelial cells, aiming at disclosing some of the mechanisms of thrombolysis of acutobin. METHODS Endothelial cells were isolated from fresh human umbilical cords by trypsin digestion of the interior surface of the umbilical vein. Cultured cells were examined by light, phase contrast and electron microscopy. The factorⅧ related antigen and CD34 of the cells were detected by AEC and DAB staining. Chromogenic assay was used to identify the activity of t PA and PAI in the medium of culture cells. Fibrin degradation products(FDPs) were measured using ELISA kit. RESULTS The cultured human umbilical endothelial cells were shown as monolayers of closely opposed, polygonal cobblestone shape by light and phase contrast microscopy. By transmission electron microscopy, cultured endothelial cells contained Weibel Palade body and showed tight junction with each other. The cells contained abundant quantities of CD34 and factorⅧ related antigen. The intercellular space among individual cell enlarged and lost polygonal cobblestone shape in the present of acutobin. Activity of t PA increased, the activity of PAI did not change significantly and FDPs increased significantly in the culture medium. CONCLUSIONS The study demonstrates the culture cells was endothelial cells according to morphologic and immunohistologic criteria. Acutobin increases the fibrinolytic activity of cultured endothelial cells and may exhibit antithrombotic effect in vivo.