Preliminary Study On Antithrombotic Mechanism Of Sea Cucumber Enzymatic Hydrolysate

Thromboembolic diseases pose a serious threat to people’s health due to their high morbidity and mortality.In recent years,studies have found that inflammation is closely related to the pathogenesis of thrombotic diseases.The inflammatory response stimulates vascular endothelial cells to release procoagulant factors such as tissue factor,which in turn activates the coagulation system,inhibits the fibrinolytic system,and eventually leads to thrombosis.At present,anti-thrombotic drugs are not effective and have certain side effects.Therefore,the development of antithrombotic drugs or supplementary foods having immunomodulatory functions from natural foods has a very bright prospect.As a kind of marine food resource with high nutritional value,sea cucumber has good anti-thrombotic activity and has been used in the development of functional foods and medicines in recent years.Previous studies have shown that the antithrombotic activity of sea cucumber is related to its polysaccharide content,and the antithrombotic effect on other active components of sea cucumber has not been reported.In this paper,the efficacy of sea cucumber enzymatic hydrolysate in antithrombotic properties was explored,and the antithrombotic mechanism was explored.This study provides a basis for the development of sea cucumbers in the prevention and treatment of functional foods or drugs for thrombotic diseases.Methods:1.Using fibrin plate to determine the fibrinolytic activity of sea cucumber enzymatic hydrolysate by quantitative and quantitative determination;2.An in vitro model of thrombus was established by in vitro natural coagulation of New Zealand rabbit blood to determine the in vitro thrombolytic activity of sea cucumber hydrolysate;3.The mice were intragastrically administrated with 350,700 and 1400 mg/kg sea cucumber enzymatic hydrolysate for 30 days,and the carrageenan model was established by carrageenan.The black tail of the mice was observed and the indicators of thrombosis were confirmed by immunohistochemistry.Thrombosis model;ELISA kit to detect fibrinolysis and coagulation indicators and inflammatory factors;4.ADP-induced platelet aggregation,qPCR analysis of tissue factor,thromboxane and prostacyclin expression.Result:1.Sea cucumber hydrolysate has good fibrinolytic activity and its fibrinolytic activity is at 0.313 U/mg;2.Sea cucumber enzymatic hydrolysate showed strong in vitro thrombolytic activity,which is in a dose-dependent manner in the range of 20-80 mg/mL;3.Through carrageenan modeling,black tail symptoms appeared in the tail of mice,which are asociated with increased FIB and VWF contents,as well as peripheral proinflammatory cytokines IL-6,IL-17,IL-23 and IFN-γ,but decreased FDP,DD,t-PA and anti-inflammatory cytokines IL-4,IL-10.The pretreatment of sea cucumbers could significantly improve the above changes.4.After ADP induced platelet aggregation,the expression of Tissue factor and thromboxane synthase was increased,and the expression rate of prostacyclin’s rate-limiting enzyme COX-2 was decreased,while sea cucumber enzymatic hydrolysate could effectively reverse the above indicators.Conclusion:1.Sea cucumber enzymatic hydrolysate has strong thrombolytic effect in vivo and in vitro;2.The thrombolytic effect of sea cucumber enzymatic hydrolysate is related to the activation of the body’s fibrinolysis system,regulation of the body’s immune balance and regulation of platelet aggregation.