少棘蜈蚣纤溶活性蛋白的抗血栓作用

Study on the antithrombotic effect of fibrinolytic proteins from Scolopendra subspinipes mutilans

目的研究少棘蜈蚣(Scolopendra subspinipes mutilans L.Koch)纤溶活性蛋白的抗血栓作用。方法采用离子交换层析和分子筛等制备法从少棘蜈蚣中纯化到蜈蚣纤溶酶(Scolopendra subspinipes mutilans L.Koch fibrinolyti cenzyme,SSFE),用纤维蛋白平板检测酶活力,常规方法检测了SSFE溶血性和出血性,进行了SSFE体外溶栓和体内溶栓实验,并检测了凝血酶原时间(PT)、活化部分凝血酶时间(APTT)和凝血酶时间(TT)3个标准化指标。结果PAGE显示SSFE为单一组分,具有纤溶活性;证明SSFE无致出血性且无致溶血活性,低、中、高剂量SSFE使小鼠APTT和TT均明显延长,中剂量SSFE对PT有延长作用,而高剂量作用不明显。结论蜈蚣纤溶酶具有抗血栓作用。

Aim The aim is to purify fibrinolytic enzyme from Scolopendra subspinipes mutilans L.Koch and to study the thrombolytic and anticoagulant effect.Methods Scolopendra subspinipes mutilans L.Koch fibrinolytic enzyme(SSFE) was purified by ammonium sulphate precipitation,DEAE-cellulose and SephadexG-75 column chromatography from Scolopendra subspinipes mutilans L.Kochby.And fibrinolytic activity was determined by fibrin plate.The anticoagulant effect was measured on mice with haemolytic test and hemorrhagic test.The thrombolytic effect was measured with rats In vitro and in vivo,and the activated partial thromboplastin time(APTT),plasma prothrombin time(PT),thrombin time(TT) were measured.Results SSFE was single component with fibrinolytic activity and without any hemolyzation and hemorrhagic activity.All doses of SSFE(2,5,10 mg·kg-1) could obviously prolong activated partial thromboplastin time(APTT) and thrombin time(TT) ;Middle dose of SSFE(5 mg·kg-1) could prolong plasma prothrombin time(PT) while high dose of SSFE(10 mg·kg-1) didn't prolong obviously.Conclusion SSFE has obvious thrombolytic effect and anticoagulant effect.