重组葡激酶-水蛭素融合蛋白在体外的抗凝溶栓作用分析  

Analysis of Anticoagulant and Fibrinolytic Function in Vitro for Recombinant Staphylokinase Hirudin Fusion Protein

作　　者：董耘[1] 徐昕[2] 李斌[1] 柯丽娜[1] 陈武[3] 

机构地区：[1]湖北医药学院附属东风医院妇产科,十堰442000 [2]湖北医药学院附属东风医院骨科,十堰442000 [3]湖北医药学院附属东风医院检验科,十堰442000

出　　处：《血栓与止血学》2015年第6期337-339,共3页Chinese Journal of Thrombosis and Hemostasis

摘　　要：目的通过构建STH融合蛋白发酵及分离纯化工艺,用以获取蛋白,并探讨其在体外的抗凝溶栓作用。方法采用琼脂糖-纤维蛋白溶圈法对样品STH的溶栓活性进行测定;SDS-PAGE、Western Blot、发色底物法以及纤维蛋白凝块溶解法分别分析STH和裂解产物的抗凝活性,并根据血栓弹力图来分析STH在体外所具有的溶栓、抗凝作用。结果在STH中,HV2的C-末端对凝血酶仍具有显著的亲和力,使STH对血栓具有较高的靶向性;STH在凝血酶裂解的情况下能将抗凝活性释放开来,是指具有溶栓、抗凝双重作用。结论 STH生产工艺操作起来简便,便于大规模生产;通过分析STH在体外所具有的功能,结果表明:STH在具有溶栓、抗凝作用的基础上,其对血栓也有显著的靶向性,但其出血副作用不大。Objective To study effect of anticoagulant and fibrinolytic function in vitro for recombinant staphylokinase hirudin fusion protein. Methods To determine thrombolytic activity of STH sample by agarose fibrin protein dissolve ring method,and to analyze anticoagulant activity of STH and the cleavage product through SDS-PAGE,Western Blot,chromogenic substrate method and fibrinclot lysis assay respectively; and evaluate thrombolysis effect in vitro according to thrombelastography. Results In STH,the C-terminal of HV2 still has an affinity to thrombin,the STH targeting to has the high thrombosis; STH in thrombin cleavage situations can be anticoagulant activity released,refers to has a dual role of thrombolysis,anticoagulation. Conclusion STH production process is simple to operate,convenient mass production; through the analysis of the STH has a function in vitro,the results showed that： STH with thrombolysis,anticoagulation based on function,it also has targetedsignificantly on thrombosis,but the bleeding side effects do not.

关 键 词：STH融合蛋白 凝血 溶栓性 

分 类 号：Q815[生物学—生物工程]