Objectives : This research was investigated the anticoagulant effect of the Gleditsiae spina extract. Methods : We researched prothrombin time (PT) assay, activated partial thromboplastin time (APTT) assay in vitro and in vivo using arteriovenous (A-V) shunt rat model and shortening Rat tail bleeding time (BT). A-V shunt and BT were treated with extract of Gleditsiae spina (GS) 400 mg/kg for a week. Results : Bleeding time of Gleditsiae spina extract in vivo had a significant increase as about 1.2 times and thrombus weight of Gleditsiae spina extract had a significant reduction of thrombus weight as 26%. Gleditsiae spina extract represented an effect of anticoagulation by operating on extrinsic pathway factor II, V, VII, X and intrinsic pathway factor VIII, IX, X, XI, XII in the coagulation system. Conclusions : Considering the above mentioned results, it is judged that a Gleditsiae Spina extract has a control effect of thrombus creation.