Research The Anticoagulant Mechanism Of Eupolyphaga Anticoagulant Component---F2-2

Objective:Eupolyphaga which is a insect drug in Tongxinluo capsule prescription, with radix paeoniae rubra and cicada slough act as adjuvant, plays the role of stasis meridians. At the early stage of our team, through the bionic enzymatic hydrolysis extraction and the dextran gel separation has got the component F2-2, which can be used as anticoagulant in vitro. And this study intends to investigate Eupolyphaga F2-2play a role in the mechanism of anticoagulant from animal experiments and cell experiments two aspects.Methods:①Normal rats were randomly assigned, to oral administration of different doses of Eupolyphaga F2-2respectively. Blooded sample from orbital venous plexus after7days, and detected t-PA and PAI-1content in plasma.②The MTT assay was used to determinate the suitable dose of eupolyphaga anticoagμlant constituent. HUVECs injury model was established after dosing, and the leakage rate of lactate dehydrogenase (LDH), superoxide dismutase(SOD)、malonal dehyde (MDA)、glutathione peroxidase (GSH-Px) and nitric oxide(NO) content of endothelial cell medium of HUVECs were measured to evaluate the protect effect of EAC.③By enzyme-linked assay for detection of the blank group and the treatment group t-PA and PAI-1levels in cell culture medium after48hours’incubation.Results:①Compared with the blank group, each dose group of F2-2t-PA and PAI-lcontent were deceased, and the higher the dosage, the lower two levels.Which in the middle and the high dose group, PAI-1levels were decreased statically different, and t-PA content reduced significantly different.②Within0～4000mg·L-1concent-ration, F2-2acted on cells24h,48h72h, the cell viability were all above80%. Compared with the control group, H2O2(37.4mg·L-1) can decrease HUVECs survival rate, which indicates successful modeling; the leakage rate of LDH and the content of MDA increased, while SOD, GSH-Px, and NO decreased significantly in the model group. However, the EAC, of which concentrations range from250to4000mg·L-1can significantly reduce LDH release and the content of MDA, increase the activity of SOD and GSH-Px comparing with the model group.③In the250～4000mg·L-1concentration range, PAI-1levels were significantly reduced in a concentration-dependent manner; but t-PA levels were increased to varying degrees, and didn’t exhibit concentration-dependent or other regularity.Conclusions:The protective effect on damaged vascular endothelial cells and a certain degree of promation effect on fibrinolytic system may be the mechanism of Eupolyphaga anticoagulant component F2-2play a role of anticoagulation.