Antithrombotic Effect Of Agkistrodon Acutus Venom By Oral Administration

Snake venom is one liquid from poisonous snake gland,the major constituent isbiologically active proteins and peptides,which is over90%to95%of the dry weight.There no report both at home or abroad about antithrombotic effect of the oraladministration of Agkistrodon acutus Venom.The task is observing the prevention andAntithrombotic effects to animal with the eon agkistrodon acutus venom and the primarytest on its system.1.Acute toxicity test of miceAcute toxicity test of KM mice was designed to calculate the median lethaldose（LD50） of the oral administration of agkistrodon acutus venom.LD50of AAV one-offig in Bliss method was640±171mg·kg^-1. LD50of rats was301mg·kg^-1on the basis ofLD50of mice in body surface area transformation method. Dose of rats and mice inaccordance with LD50.2.Anticoagulant effects of AAV on APTT、PT and TTAnticoagulant effects were determine by activated partial thromboplastintime（APTT）、prothrombin time（PT） and thrombin time（TT）.Both AAV40and80mg·kg^-1ig could both significantly increase APTT. The timing of making APTT by AAV40、80mg·kg^-1were48.8±12.79s and42.0±8.40s, with obvious difference comparedwith NS（P<0.01,P<0.05）.There was no statistics meaning to PT and TT.It can beseen AAV inhibits intrinsic coagulation pathway.3.Protective effect of AAV on mice with pulmonary thrombus induced by ADP-Na2We observed the effect on the respiratory distress due to pulmonary thrombosisinduced by ADP-Na2after AAV60、120mg·kg^-1ig were28.00±6.07s and37.38±6.09s respectively, asprin was45.75±9.88s,with obvious difference compared withNS（P<0.01,P<0.05）.AAV markedly relieved the respiratory distress.4.Effect of AAV on thrombolysis of rats in man-made artery-vein circulationThe effect of AAV was estimated by weight of thrombus. AAV40、80mg·kg^-1igcould significantly reduce the weight of thrombus. Comparing with the thrombus weight,AAV40、80mg·kg^-1were31.98±1.04mg and27.48±0.77mg respectively, asprin was28.13±2.76mg,with obvious difference compared with NS（P<0.01）.AAV has theinhibition to thrombosis of rats common carotid artery in man-made artery-veincirculation. 5.Thrombolytic effect of AAV on thrombus of inferior caval vein induced by powerof thrombinThe thrombus of inferior caval vein was induced by power of thrombin.AAV40、80mg·kg^-1ig could significantly ruduce the weight of thrombus. Comparing with thethrombus weight,AAV40、80mg·kg^-1were55.08±2.56mg and35.53±2.52mgrespectively, lumbrukinase was34.36±4.22mg，with obvious difference compared withNS（P<0.01）.AAV has the inhibition to thrombosis of rats inferior caval vein induced bypower of thrombin.6.Antithrombotic effect of AAV on mice with caudal vein thrombosis induced bycarrageeninThe effect of AAV was estimated by the length of infarcted regions. AAV40、80mg·kg^-1ig could significantly reduce the length of infracted regions. Comparing with thelength of infarcted regions, AAV40、80mg·kg^-1were31.98±1.04mg and27.48±0.77mg respectively, asprin was28.13±2.76mg,with obvious difference compared withNS（P<0.01）.AAV has the prevention and inhibition to thrombosis of mice in caudal vein.7.The effect on euglobulin lysis time after AAV oral administration of ratsThe effect of AAV was estimated by euglobulin lysis time of rats in plasma. AAV40、80mg·kg^-1ig could significantly reduce the euglobulin lysis time. Comparing withthe euglobulin lysis time, AAV40、80mg·kg^-1were342±34.2min and360±56.1minrespectively, with obvious difference compared with NS（P<0.01）.8.The changes of t-PA content after AAV oral administration of ratsThe mechanism of AAV antithrombotic effect was estimated by t-PA content of ratsin plasma. AAV40、80mg·kg^-1ig could significantly increase the t-PA content.Comparing with the t-PA content, AAV40、80mg·kg^-1were207.4±46.38pg·ml^-1and375.6±97.92pg·ml^-1respectively, with obvious difference compared with NS（P<0.01）.9.Fibrinolytic activity of AAVSR in vitroAAVSR fibrinolytic activity was determined by fibrin plate process. AAVSR of40、80mg·kg^-1ig were obviously fibrinolytic. Comparing with the dissolve circle area, AAV40、80mg·kg^-1were64.97mm2and98.81mm2respectively, Lumbrokinase was78.12mm2,with obvious broader than NS（2.91mm2）.AAV also had effect on fibrin platewithout fibrinogen.AAV exhibit indirect and direct fibrinolytic effect.10.The effect on blood-clot lysis time of AAVSR in vitroAAVSR of40、80mg·kg^-1ig could significantly reduce coagula weight. Comparingwith the thrombolysis rate, AAV40、80mg·kg^-1were19.23±0.05and21.27±0.02respectively, with obvious difference compared with NS（P<0.05,P<0.01）. ConclusionAAV40、80mg·kg^-1ig could significantly increase activated partial thromboplastintime of rats plasma,shorten euglobulin lysis time,increase t-PA content;AAV40、80mg·kg^-1ig have the inhibition to thrombosis of rats common carotid artery in man-madeartery-vein circulation;AAV40、80mg·kg^-1ig have the inhibition to thrombosis of ratsinferior caval vein induced by power of thrombin;AAV60、120mg·kg^-1ig have theinhibition and lysis to thrombosis of mice in caudal vein;AAV60、120mg·kg^-1ig relievethe respiratory distress;AAVSR had indirect and direct fibrinolytic activity;AAVSR couldsignificantly increase thrombolysis rate;there is dose-relationship treated with AAV.