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本文报道自尖吻蝮蛇中分离、纯化三个凝血酶样酶(Thrombin-like Enzymes,TLEs),TLE-A,TLE-Ⅵ及TLE-Ⅶ。双向免疫扩散分别出现单一沉淀线,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分别呈单一区带,并依此测定分子量分别为66、31及14KD。试管内TLES能使纤维蛋白原或血浆形成纤维强白微凝块。凝血酶样酶制剂(TLES)150μg/ml效价与牛凝血酶1.0U/ml接近。肝素不能对抗其凝血效应,以对甲苯磺酰精氨酸甲基酯(TAME)为底物,其精氨酸酯酰活力可达2000μmol/min·mg左右。动物在体实验中,犬(n=6)静脉注射100μg/kg后4h药效达高蜂,血浆纤维蛋白原含量明显降低(P<0.02),凝血酶时间明显延长,血浆副凝血试验强阳性,血小板聚集功能明显抑制,家兔(n=10)每日静脉注射300μg/kg连用7天,对血液凝固系统效应与犬相似。药后第8天处死家兔病理解剖无明显变化。小鼠一次静脉注射的LD_(50)及其95％可信限为14.5±2.2(12.3～16.6)mg/kg。
In this paper, three Thrombin-like Enzymes (TLEs), TLE-A, TLE-Ⅵ and TLE-Ⅶ were isolated and purified from Agkistrodon acutus. Two-dimensional immunodiffusion showed a single precipitation line, respectively. SDS-PAGE showed a single zone, and the molecular weights were 66, 31 and 14 kD, respectively. In-vitro TLES enables fibrinogen or plasma to form fibrillar micro-clots. Thrombin-like enzyme preparation (TLES) 150μg / ml titer and bovine thrombin 1.0U / ml close. Heparin can not counteract its clotting effect, and its arginine ester acyl activity can reach about 2000μmol / min · mg with TAME as substrate. Animals in vivo experiments, dogs (n = 6) intravenous injection of 100μg / kg 4h after the effect reached a high bee, plasma fibrinogen levels were significantly lower (P <0.02), thrombin time was significantly longer, plasma paranex test was strongly positive , Platelet aggregation function was significantly inhibited, rabbits (n = 10) daily intravenous injection of 300μg / kg for 7 days, the effect of blood coagulation system similar to dogs. The rabbits were sacrificed on day 8 after the pathological anatomy had no significant change. The LD_ (50) and its 95% confidence limit for a single intravenous injection in mice were 14.5 ± 2.2 (12.3-16.6) mg / kg.