蝮蛇毒血小板抑制因子对动脉血栓形成的影响及机制研究

Effect and mechanism of platelet inhibitor from Agkistrodon halys venom on artery thrombosis

目的:探讨皖南蝮蛇毒血小板抑制因子(AHV-PI)对家兔动脉血栓形成的影响及可能机制。方法:新西兰家兔24只,随机分成假手术组、动脉血栓模型组、阳性对照组(奥扎格雷钠,5mg/kg),AHV-PI(0.1mg/kg)实验组共4组,每组6只。应用70%FeCl3溶液化学损伤的方法来制备家兔颈动脉血栓模型,采用血栓弹力仪(TEG)描计血栓弹力图,比浊法测定家兔血小板聚集率,ELISA测定各组血浆中α颗粒膜蛋白(GMP-140)和血栓素B2(TXB2)水平,光镜和透射电镜分别观察动脉血栓形成和血小板形态的改变。结果:AHV-PI实验组与模型组相比,血栓弹力图凝血时间(R)值和血凝块形成时间(K)值延长(P<0.01和P<0.05),Alpha角度、最大幅度(MA)和凝血指数(CI)减小(P<0.05和P<0.01);血小板聚集率的各项指标均明显降低(P<0.05);血浆中GMP-140和TXB2含量降低(P<0.01)。AHV-PI实验组光镜下动脉内未见血栓形成,血小板电镜显示血小板形态基本规则,与模型组相比伪足较少,α-颗粒和致密颗粒无明显减少,胞浆空泡化现象减轻。结论:AHV-PI可以通过抑制血小板聚集,防止动脉血栓的形成,其机制可能与之保护血小板超微结构,减少血小板脂质代谢和颗粒内容物的释放有关。

AIM： To explore the effect and mechanism of platelet inhibitor from Agkistrodon halys venom（AHV-PI） on artery thrombosis in rabbits.METHODS： Twenty-four New Zealand rabbits were randomly divided into normal control,artery thrombosis model,positive control（sodium ozagrel at dose of 5 mg/kg） and AHV-PI（0.1 mg/kg） experiment groups（n=6 for each）.The model of rabbits carotid artery thrombosis was induced by infiltering 70% FeCl3.Thrombelastogram was obtained by thromboelastography instrument.The experiment of platelet aggregation was determined by the turbidimetric method.The content of GMP-140 and TXB2 in plasma were determined by ELISA.The structural changes of the artery thrombosis and platelet were observed under microscope and transmission electron microscope respectively.RESULTS：Compared with model group,R and K were significantly prolonged in AHV-PI experiment group.The alpha angle,MA,CI and the indices of platelet aggregation were decreased（P0.01 and P0.05）.The levels of GMP-140 and TXB2 in plasma were decreased（P0.01）.Artery thrombosis histological observation showed that thrombosis in AHV-PI experiment group were not found.Platelet electron microscope showed that the platelet morphology and ultrastructure approach normal control group.AHV-PI can inhibit producing pseudopods process and increase in the number of theαgranules associated with enhanced intensities of their electron densities compared with model group.CONCLUSION： AHV-PI can inhibit platelet aggregation and provent the information of artery thrombosis.The mechanism may be attributed to protecting platelet ultrastructure and reducing the release of platelet lipid metabolism and particulate contents.