Coagulant Effects and Mechanism of (L.) Frodin.

(L.) Frodin, are commonly used in anti-inflammatory, analgesic, traumatic bleeding and hemostasisas. In this paper, the coagulation effect of the ethanol extract (Set), ethyl acetate phase (Sea) and -butanol phase (Sbu) was evaluated by prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and fibrinogen content (FIB) assays in vitro. Then, Three main lupanine triterpenes (compounds -) were isolated and identified from Sea and Sbu by a bioassay-guided method and their structure were identified as 3α-Hydroxy-lup-20(29)-ene-23, 28-dioic acid, betulinic acid 3--sulfate and 3α-Hydroxy-lup-20(29)-ene-23, 28-dioic acid 28--(α-l-rhamnopyranosyl(1→4)--β-d-glucopyranosyl(1→6))-β-d-glucopyranoside) by spectroscopic data analysis. Among of them, compound was confirmed to have significant coagulant effect in vitro. Furthermore, the pro-coagulation mechanism of extracts and compound were investigated by measuring whole blood viscosity (WBV), plasma viscosity (PV), erythrocyte sedimentetion rate (ESR), pack cell volume (PCV), APTT, PT, TT, and FIB in vivo. Meanwhile, the levels of thromboxane B (TXB), 6-keto prostaglandin F (6-keto-PGF), endothelial nitric oxide synthase (eNOS) and (endothelin-1) ET-1 were detected. The bleeding time (BT) was tested by tail bleeding method, which proved the traumatic bleeding and hemostasis activities of . The pharmacology experiments showed that the Set, Sea, Sbu and compound has significant pro-coagulation effect. In addition, compound might be the main constituent of pro-coagulation in These results could support the fact that could be used traditionally to cure traumatic bleeding, and the pro-coagulation effects were associated with the regulation of vascular endothelium active substance and hemorheology parameters.