Screening Of Potential Anti-thrombotic Ingredients From Salvia Miltiorrhiza In Zebrafish And By Molecular Docking

Thrombosis can directly or indirectly cause the occurrence and development of a series of cardiovascular and cerebrovascular diseases(CVDs).Since the occurrence of CVDs can be reduced by the inhibition of thrombosis,study of new antithrombotic drugs has been a hotspot all the time.In the theory of traditional Chinese medicine(TCM),promoting blood circulation and removing blood stasis is the key to the treatment of thrombosis.Salvia miltiorrhiza Bge.(Danshen,DS)is one of the most frequently used drugs in TCM,treating for the patients with blood circulation problem.However,the active ingredients in DS and the mechanism of action is still unclear.In our study,molecular docking technology and zebrafish in vivo thrombosis model was used to screening the antithrombotic active ingredients in DS.The results were obtained as the follows:Screening of potential antithrombotic active ingredients in DS.Salvianolic acid B(SAB),rosmarinic acid(MA),lithospermic acid(LA)and luteolin-7-O-β-D-glucoside(LG)in DS that may have potential antithrombotic effects were screened out with molecular docking technology.All of the four compounds exerted antithrombotic effects through multiple targets.The zebrafish thrombosis model was constructed and optimized.In the experiment,two chemical inducers,(±)-Eplnephrlne hydrochlorlde(AH)and Phenylhydrazine(PHZ),were used to construct two stable and repeatable zebrafish thrombosis models,respectively.The relevant optimized parameters for model constructed were: Zebrafish at 3 dpf(days post-fertilization)treated with AH at a concentration of 15.0 μM for a time period of 16 hours and then stained with O-dianisidine staining solution for 30 min;Zebrafish at 3 dpf treated with PHZ at a concentration of 1.5 μM for a time period of 24 hours and then stained with O-dianisidine staining solution for 30 minThe antithrombotic effects of SAB,MA,LA and LG were proved.The result of our study showed that the active components SAB,MA,LA and LG in DS could exert antithrombotic effects before or after thrombosis induced by chemical inducers AH and PHZ in zebrafish.This result was coincident with the conclusion of molecular docking to a certain extent.