Study On Purification, Characterization And Thrombolytic Effects Of Lumbrokinas

The experiment was conducted to achieve an effective method of purifying lumbrokinas (LK) from Eisenia fetida earthworm and study the characteristics of LK and the thrombolytic effect of LK.Expt.1: Purification and Characterization of Lumbrokinase LK was purified through extraction,(NH4)2SO4 fractional salting out,Sephadex G-75,DEAE-Sepharose 6 Fast Flow and Phenyl Sepharose High perfermance from earthworm Eisenia foelide and identified by SDS-PAGE.Effects of pH value, temperature and inhibitions on the activity of LK and the mechanism of LK were studied . The specific activity of LK amounts to 5 100 U/ mg. Two fractions of LK, with Mr of 32 000 and 33 500, was obtained through SDS-PAGE. LK kept high activity in neutral and leaning alkalinity environments.LK activity was stable in wildly temperature.respectively; The effects of different types and concentrations of metal ions on the activity of LK were different.Studies on substrate specificity and inhibition indicated that LK belonged to serine protease but metalloproteinase, and had disulfide bonds. Purified LK can be obtained by the the techniques of the separation and purification. Purified LK can be obtained by the techniques of the separation and purification. The fibrinolytic experiment in vitro indicated that LK not only hydrolyzed fibrin directly,but also activated the plasminogen to plasmin Fibrinolytic enzyme.Expt.2: Effect of Lumbrokinase on thrombosis and coagulation indexs induced by carrageenan in rats.The experiment was conducted to evaluate the thrombolytic effect of LK on rats induced by carrageenan.The results shows that: the high dose LK can obivous decrease the black tail occurrence and the black tail length (P<0.05),weaken the lengthening of the black tail. Both the low LK (P>0.05) and the high dose LK (P<0.05) can lengthen the PT, TT and fibrinogen; the high dose LK also can weaken the lose of the body weight induced by carrageenan. In a word, LK could improve the body fibrinolytic function, prevent and relieve the thrombosis.