皖南蝮蛇毒PCA活化血浆蛋白C的特性研究

Protein C activator from the venom of agkistrodon halys for activation of plasma protein C

目的:研究皖南蝮蛇毒蛋白C激活物(protein C activator,PCA)活化血浆蛋白C(protein C,PC)的特性。方法:家兔待检血浆分别与不同浓度的PCA混合后,行PT及APTT测定;随后利用PC活性检测试剂盒,检测不同浓度PCA对PC活性的影响;最后将最适浓度PCA分为PCA组、PCA血浆组、PCA血浆加PC抗体组,并应用发色底物法配合酶标仪对各组活化PC的能力进行鉴定。结果:与正常血浆相比,低剂量PCA即可显著延长APTT(P<0.01),随着PCA浓度的增加PT也显著延长(P<0.01);经检测0.3 mg/ml浓度的PCA具有最适PC活化特性,与阳性对照Protac量效相当(P>0.05))。结论:皖南蝮蛇毒PCA具有与Protac效应相当的活化PC特性,提示蛇毒PCA具有一定的临床应用前景。

Objective：To examine the property by using protein C activator（PCA） from the venom of agkistrodon halys from south Anhui province to activate plasma protein C（PC）.Methods：The plasma from rabbits was mixed with PCA of different concentration and determined for prothrombin time（PT） and activated partial thromboplastin time（APTT）,followed by examining the influence of PCA in separate concentration on PC with PC Activity Assay Kit.Subsequently,the PCAs with optimal concentration were allocated to groups of simple PCA,PCA plasma and PCA plasma plus PC antibody for determining the competency of PCA in each group to activate PC by chromogenic substrate with microplate reader（model 680）.Results：Compared with the normal plasma,low concentration of PCA could significantly delay APTT（P〈0.01）,and yet,PT was also delayed markedly with increased concentration of PCA（P〈0.01）.The determination showed that PCA in concentration of 0.3 mg/ml presented with excellent performance to activate PC,which was consistent with the dose-response relationship by positive control of Protac.Conclusion：The PCA isolated from the venom of agkistrodon halys from south Anhui province appears as competently as Protac in activating PC,suggesting that this venom has clinical applications.