Effects Of Scorpion Purified Liquid On The Release Of Cultured Vascular Endothelia Cells Induced By Thrombin

Objective:The human umbilical vein vascular endothelia cells (HUVECs) cultured in vitro were selected for the study,to observe the effect of scorpion purified liquid(PLIS) on the release of HUVECs induced by thrombin,and to investigate the mechanism of anticoagulation.Methods:The HUVECs was cultured in vitro, the well grown 2-3 generation HUVECs were divided into control group(1640 culture medium),high concentration PLIS group (PLIS 24μg/mL+thrombi n 10U/mL),medium concentration PLIS group(PLIS 12μg/mL+thrombin 10U/mL),low concentration PLIS group(PLIS 6μg/mL+thrombin 10U/mL). After HUVECs were cultured for 12 hours the cell morphology was observed,the supernatant and Cell lysates was collected in each group.①The concentration of von willebrand factor (vWF), 6-keto-prostaglandin F1α(6-keto-pGF1α), tissue plasminogen activator(t-PA), and plasminogen activator inkibitor (PAI-1) were measured with enzyme linked immunosorbent assay (ELISA);②The activity of tissue Factor (TF), tissue factor pathway inhibitor (TFPI) were measured with Chromophore substrate method. Results:1. Compared with the blank control group, morphology of HUVEC cells in thrombin group were changed,the PLIS in all concentration inhibited the effects of thrombin. The experimental results demonstrate that PLIS can protective HUVEC induced by thrombin.2. The concentration of vWF was significantly increased in thrombin group, Compared with blank control group P<0.01;The concentration of vWF was obviously decreased in high,medium,low concentration PLIS groups, Compared with thrombin group all P<0.01.3. PLIS is able to maintain the dynamic balance of fibrinolytic system and to balance the disturbance of fibrinolysis coagulation system induced by thrombin.The concentration of PAI was decreased and tPA was increased in high,medium,low concentration PLIS groups,compared with thrombin group the difference was significant(p<0.01).4.6-keto-pGF1αwhich is inactive metabolites of PGI2 has the function of dilatesing blood vessels, inhibiting platelet aggregation, stabilizeing membranes. The concentration of 6-keto-pGF1αwas decreased in thrombin group(compared with blank control group) but increased in high,medium,low concentration PLIS groups(compared with thrombin group). The results show that PLIS has the effects of inhibiting platelet aggregation.5. PLIS isableto inhibit TF express but has no influence to the release of TFPI.Conclusion:PLIS has the obvious effect of anticoagulation, the mechanism of anti coagulation similar to hirudin. through inhibit coagulation pathway to play a part in anticoagulation...