Study On The Anticoagulant Effect And Mechanism Of The Total Saponins Of Polygala Fallax Hemsl Based On Network Pharmacology

Objective: This study aims to predict the anticoagulant target and mechanism of the total saponins of Polygala fallax Hemsl by means of network pharmacology.The mechanism of the predicted target is further verified.The predicted target and mechanism are further verified by the in vitro anticoagulation index detection experiments and animal experiments.Methods: Ethanol hot reflux method was used to extract PTS(total saponin of Polygala fallax Hemsl);HPLC(high-performance liquid chromatography)was used to determine the content of saponin A in PFH(Polygala fallax Hemsl),and the content of PTS was determined by UV(ultraviolet spectrophotometry).All the chemical components and targets of PFH were collected through the means of network pharmacology,and targets and pathways of PTS were screened and predicted.Fe Cl3 induced rat carotid artery thrombosis model test verifies the anticoagulation and antithrombotic effects of PTS.It determines the classical in vitro anticoagulation parameters APTT(activated partial thromboplastin time),TT(thrombin time),and the effect of PT(Prothrombin time)activity and the effect of blood coagulation factor/cofactor-FIIa and FXa activity,which initially determine its anticoagulant mechanism.Results: the content of PFH Saponin A in this batch of PTS was 1.673%,the total saponin content was 37.091%.The anticoagulant targets of PTS predicted by network pharmacology were FII(coagulation factor II)and PTAFR(Platelet Activating Factor Receptor).The animal experiments show that PTS can significantly inhibit the formation of Fe CI3 induced rat carotid artery thrombosis,the inhibition rate of the high-dose PTS group was 42.68%,and the ratio of thrombosis area is 43.51%.In vitro anticoagulation experiments show that PTS can significantly prolong the quality control plasma APTT and TT in a dose-dependent manner.It has no effect on PT,The required concentration for doubling human control plasma APTT and TT is 0.36mg/m L and 0.14mg/m L,respectively in the range of 0-2mg/m L,which indicates that PTS exerts its anticoagulant effect by affecting the endogenous coagulation pathway and the common coagulation pathway.Blood coagulation factor/ cofactor detection experiments further revealed that PTS exerts its anticoagulant effect by inhibiting the FIIa,and that is thrombin activity.Its IC50 is85.88±12.50?g/m L.Conclusion: In vivo and in vitro experiments confirmed that the total saponins of PFH had anticoagulant effect,and its anticoagulant mechanism was to inhibit FIIa activity.The pharmacological analysis and prediction based on the component target network are basically consistent with the experimental results.It mainly works on the activated FII.