Parametric Study Of The Antithrombotic Activity Of Protease In Pheretima

The Pheretima aspergillum(E.Perrier ), also customly named Guang earth-worm, belongs to the family of Megascolecidae. Earth-worm is a frequently used Chinese materia medica . In Chinese Pharmacopeia ,three other kinds of species of earth-worms are recorded ,including P. Vulgaris chen , P. Guillelmi(michaelsem) and P. pectinifera michaelsen , customly named Hu earth-worm. Property of earth-worm is cold ; taste is salty ; Belong to liver.spleen and bladder channels. The function is removing heat, calming wind, dredging meridian, pacifying gasp, diuresis and heat-ache, etc.It showed that Pheretima aspergilIum(E.Perrier ) and Eisenia foetida had Thrombolytic activity and there was not much different Thrombolytic activity between them through the activity contrast study between two kinds of earth-worm . It showed that there were proteins with close molecular weight both of them by electrophoresis and HPLC. It showed that there the close relationship between them through the Random Amplified Polymorphic DNA (RAPD). All of clinical drug from earthworm are crude product.Thrombolytic enzyme from Pheretima aspergilIum(E.Perrier ) was studied in this paper . 5 kinds of proteins from guang-earthworm were purified by salt out , film , CM-sepharose , DEAE-sephadex-A50, sephacryl-S200 and sepharose F.R.one of them have high Thrombolytic activity. And their molecular weight are >97000,60500,47900,28100 and 26300. Amino acidic components of the 5 kinds of proteins were analyzed by amino acidic automatic analysis instrument. One of proteins was determined by biosystem process precise ,it is Gln-Iie-GIy-Gly-Thr-Asn-Ala-Ser-Pro-Gly-Glu-Phe-Pro-Pro-Gln -Leu-Ser-Gln-Thr.Quality standard of Pheretima aspergillum(E.Perrier ) was determined . Extracting process was Optimized according to the results of the orthogonal experiment, and the Quality standard of the extract of guang-earthworm was determined including activity and fingerprint . Thrombolytic activity of extract was confirmed through model of A-V bypass.