蕲蛇酶、凝血酶对人及牛纤维蛋白原的凝血反应比较

Comparison of Clotting Effects of Acutobin and Thrombin on Human and Bovine Fibrinogens

目的 研究蕲蛇酶、凝血酶与不同种属来源的纤维蛋白原的凝血作用的差异 ,以选择检定蕲蛇酶活力的最适底物。 方法 取不同浓度人凝血酶与人纤维蛋白原 (HFg)、牛纤维蛋白原 (BFg)及人血小板血浆 (PPP)反应 (试管法或用血液凝聚仪 ) ,分别记录初凝时间并求反应曲线的回归方程。蕲蛇酶也稀释成不同浓度依上法测定 ,并求回归方程。另以人凝血酶 (1 .2 5IU/ ml)、蕲蛇酶 (1 .2 5AU/ ml)与梯度浓度的 HFg反应并求最大反应速度。此外还测定了蕲蛇酶以家兔 PPP为底物的凝血活力和以 BAEE、TAME为底物的精氨酸酯酶活力。 结果 人凝血酶、蕲蛇酶对 HFg的反应速度均比 BFg快 (P <0 .0 5) ;丝氨酸蛋白酶抑制剂 PMSF和金属蛋白酶抑制剂 EDTA均能抑制蕲蛇酶的凝血活力 ,但后者不能抑制蕲蛇酶的精氨酸酯酶活力。

Objective To study the clotting effects of acutobin and thrombin on various sources of fibrinogens to select optimal substrate for assay the activity of acutobin Methods Various concentration of human thrombin or acutobin were reacted with human, bovine fibrinogen or human platelet poor plasma (PPP) in 37℃ water bath and the clotting time were recorded respectively The equation of linear regression were obtained Another test was the maximal velocity of human thrombin (l25 IU/ml) and acutobin (l25AU/ml) with gradient concentration of human fibrinogen The clotting time with rabbit's PPP and the arginine ester hydrolase activity of acutobin were also assayed Results The clotting time of thrombin or acutobin with human fibrinogen were shorter than with bovine fibrinogen (P<005) The clotting activity of acutobin was inhibited by EDTA and PMSF (an inhibitor of serine protease)But its arginine ester hydrolase activity was only inhibited by PMSF Conclusion The clotting reaction of acutobin and thrombin with human fibrinogen were stronger than with bovine fibrinogen It seems that the human fibrinogen is a more suitable substrate for acutobin activity assay