Comparative Study on Anticoagulant Activity in Vitro of Two Kinds of Pheretima by Different Extraction Methods

OBJECTIVE To study the effect of wine processing on the anticoagulant activity of Pheretima in vitro, and to provide scientific basis for clinical application of Pheretima. METHODS The traditional water extraction method and bionic enzymatic extraction method were used to extract the raw and wine made Pheretima. Activated partial thromboplastin time(APTT), prothrombin time(PT), thrombin time(TT) and antithrombin activity were seleced as activity indexes to evaluate the anticoagulant activity of raw and wine made Pheretima. Then BCA method was applied to measure the content of soluble protein and polypeptide. RESULTS When water extraction was used, the results of PT and antithrombin titration showed that the anticoagulant activity of raw products and wine-processed products was the same. The results of TT showed that wine-processed products had stronger anticoagulant activity than raw products. The results of APTT showed that Pheretima had procoagulant activity, and in a certain range, the higher the concentration, the stronger the procoagulant effect. The content of protein and polypeptide in wine-processed products was higher than that in raw products. When biomimetic enzymolysis extraction method was used, the results of APTT and PT showed that the anticoagulant activity of wine-processed products was stronger than raw products. The results of protein and polypeptide content showed that the content of wine-processed products was higher than that of raw products. TT method and antithrombin titration method were not compared the difference between wine-processed products and raw products. CONCLUSION Compared with the traditional water extraction method, the bionic enzymatic extraction method can extract more protein and peptide components, and the bionic enzymatic hydrolysate has stronger anticoagulant activity. The results of the two extraction methods show that the wine making of Pheretima is beneficial to the dissolution of soluble protein and polypeptide, and could enhance the anticoagulant activity in vitro.