Studies On The Thrombolytic Effect Of Fibrinolytic Enzyme-rich Douchi Vacuum Freeze-dried Powder

Douchi is a traditional Chinese fermented soybean product used as food and medicine since ancient times,but the product is just generally used as a condiment.Fully study and excavation of the effect of Douchi and Douchi fibrinolytic enzyme is of great significance for the development of Douchi,but few researches was done in this area home and abroad.To build a foundation for further study,this paper investigates the safety and in vitro and in vivo thrombolytic effect of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder made by our laboratory.To study the in vitro thrombolytic effect of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder, in vitro blood clots dissolving experiments were conducted.The results showed that,20.0% Douchi extract can dissolve in vitro thrombus with dissolving rate 95.07±0.86%.In another experiment,the treated group,the fibrinolytic enzyme of which was inactivated by heating,cannot dissolve in vitro thrombus,with dissolving rate12.48±2.43%.It indicates that the thrombolytic substance in the powder is Douchi fibrinolytic enzyme or heat-sensible micromolecule.Similarly,in the in vitro blood clots dissolving experiment of Douchi fibrinolytic enzyme concentrate,each dose of the concentrate,the micromolecule of which was removed by dialysis,has the in vitro thrombolytic effect,whereas the treated group don't.Therefore,the fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder has in vitro thrombolytic effect,with Douchi fibrinolytic enzyme as its thrombolytic substance.As a prerequisite for in vivo thrombolysis study,test of maximum tolerated dose was introduced to assess the acute toxicity of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder.The results showed that,no mouse died within 14 days and non-lethal features were not affected by the powder.The powder's oral acute toxicity dose is greater than 16g/kg·bw,belonging to the non-toxic level.From this, the safety of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder for human is preliminary determined.In order to study the in vivo effect of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder,its effect on coagulation system in normal animals was determined.The results showed that, fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder has no effect on APTT and PT,but can prolong TT.It reveals that,the powder affects anticoagulation system or fibrinolysis system without affecting endogenous and exogenous coagulation systems in normal rats.In addition,rat abdominal aortic thrombosis model was constructed to explore the effect of fibrinolytic enzyme-rich Douchi extract on the model.The results showed that, fibrinolytic enzyme-rich Douchi extract can reduce thrombus weight,prolong the thrombin time,increase fibrin(ogen) degraded products,without affecting plasma fibrinogen concentration,and pathological section reveals that,the extent of blood vessel blocking by thrombus can be reduced by the extract.Therefore,the fibrinolytic enzyme-rich Douchi extract has thrombolytic effect in vivo,it can hydrolysis fibrin without causing bleeding side effect.Therefore,the safety and in vitro and in vivo effects of fibrinolytic enzyme-rich Douchi vacuum freeze-dried powder are determined,and the powder desearves further study.