几种水蛭抗凝血物质提取及活性分析

Anticoagulant Active Substances Extraction and Anti-Thrombin Activity Analysis of Several Species of Leeches

研究3种提取方法对菲牛蛭不同处理品的抗凝作用,采用筛选的最佳提取方法分析了5种水蛭各处理品的抗凝血酶活性,并比较了3个不同地理分布野生菲牛蛭和2个不同饵料饲喂菲牛蛭各处理品的抗凝血酶活性。结果表明:丙酮浸提法在菲牛蛭各处理品中均具有最高的提取得率,所测抗凝血酶活性显著高于其他方法(P<0.05);5种水蛭抗凝血酶活性测定结果显示非吸血蛭类的湿体匀浆液和干体无或微弱的抗凝活性,而吸血蛭类(日本医蛭和菲牛蛭)具有较高抗凝活性,且菲牛蛭的抗凝活性显著高于日本医蛭(P<0.01);不同地理分布野生菲牛蛭和不同饵料饲养菲牛蛭的湿体匀浆液、干体粉末和唾液腺分泌物抗凝活性均无显著差异(P>0.05)。研究表明菲牛蛭具有较强的体外抗凝血酶活性,且人工养殖对菲牛蛭的抗凝活性未产生不利影响。

The anticoagulant activity in vitro of three different sample isolated from the leech, Hirudinaria manillensis with three different extracting methods was studied, and utilized the optimal extracting process, the anti-thrombin bioactivity of five species of leeches were analyzed, and compared the anti- thrombin bioactivity of three species of wild H. manillensis from different geographical distribution and two species of cultured H. manillensis reared with different diets. The results showed that among them, the extracts produced by acetone precipitation had the highest yield, and also displayed the stronger anti- coagulant activity tested than others （P 〈 0.05 ）. The test displayed three species of non-bloodsucking leeches （Whitmania pigra, Whitmania acranulata and Whitmania laevis） had no or lower anti-thrombin activity, and the two species of blood-sucking leeches （H. manillensis and Hirudo nipponia） had higher anti-thrombin activity, and that the anti-thrombin activity of H. manillensis was higher than Hirudo nip- ponia significantly （ P 〈 0. 01 ）. There was no significant difference of inhibitory activity to thrombin be- tween wild and cultured H. manillensis （P 〉 0.05 ）. In conclusion, the leech of H. manillensis had high anti-thrombin bioactivity in vitro, and the artificial breeding had no adverse effect on its quality and anti-coagulant activity.