Effect Of Bombyx Batryticatus Injection On The Thrombosis’ Mechanism Of Human Umbilical Vascular Endothelial Cell Induced By Thrombin

Objective:To observe the effect of bombyx batryticatus injection on the tissue type plasminogen activator (t-PA)、plasminogen activator inhibitor (PAI)、prostacyclin(PGI2)、von willebrand factor(vWF) and tissue factor(TF)、tissue factor inhibitor(TFPI) activities of human umbilical vascular endothelial cell induced by thrombin.Methods:MTT method is according to the succinic acid dehydrogenase in the living cell line plastochondria’s which can cause the exogenetic MTT return to original state of the soluble royal purple crystal, and deposits in the cells, this crystal can dissolve in the organic solvent. Its formation is directly proportional to the amount of living cells with and often uses to reflect the cell multiplication indirectly activeness. It can use MTT method to examine cytotoxin function of the thrombin and the bombyx batryticatus injection with the HUVEC, then take next step tests with the cytotoxin active medicine density(suppression rate) which is smaller than 20% the work density. The well cultured 2-5 generation HUVEC for the study were divided randomly into common control group、thrombin group、small concentration of bombyx batryticatus injection、middle concentration of bombyx batryticatus injection、large concentration of bombyx batryticatus injection and incubated for 24 hours after various kinds of factors function.1、To observe morphology of the cells; 2、To observe the activities of PAI and t-PA; 3、To observe the content of vWF and the metabolite of PGI2 (6-Keto-PGF1α) released with ELISA method; 4、To observe the TF and TFPI activities.Results:1、After the process of pasting the wall, the extension, the fission to multiply, normal endothelial cells are covered with the entire board obviously under the low time of microscope and formated "the paving stone" crustification arrangement. Incubation with thrombin decreased the amount of cultured cells and changed their morphology, the partial cells may occur fall off and present the unusual pellets when the thrombin density is 20U/ml. Cytotoxin activeness is smaller than 20% when the thrombin density is 10U/ml, Cytotoxin activeness is smaller than 20% when the bombyx batryticatus injection density is 600μg/ml,so the thrombin density is 10U/ml and bombyx batryticatus injection density is 600μg/ml finally, then small concentration of bombyx batryticatus injection、middle concentration of bombyx batryticatus injection、large concentration of bombyx batryticatus injection are respectly 600μg/ml、300μg/ml and 150μg/ml according to the 4:2:1 proportion; 2、thrombin promoted the activities of t-PA、PAI respondly (compared with the common controls P<0.05), but the activities of t-PA were increased and the activities of PAI were reduced by small、middle and large concentrations of bombyx batryticatus injection respectly (compared with thrombin group P< 0.05); 3、thrombin promoted the content of vWF and 6-Keto-PGF1α(compared with the common controls P< 0.05), but the content of vWF were reduced by small、middle and large concentrations of bombyx batryticatus injection respectly (compared with thrombin group P<0.05), but the content of 6-Keto-PGF1αwere not effected(P>0.05);4、the activities of TF and TFPI were increased after stimulated by thrombin, the activities of TF with small、middle and large concentrations of bombyx batryticatus injection all reduced(compared with thrombin group P<0.05), but the TFPI were not effected(P>0.05).Conclusion:Bombyx batryticatus injection can significantly inhibit the HUVEC releasing function induced by thrombin and inhibit the coagulation process.