rAcAP5的溶栓作用及作用机制研究

doi:10.5246/jcps.2012.01.009

rAcAP5的溶栓作用及作用机制研究

丁双, 刘晓岩, 朱元军, 王银叶^*

北京大学医学部药学院分子与细胞药理学系, 北京 100191

收稿日期:2011-05-09 修回日期:2011-09-20 出版日期:2012-01-01 发布日期:2012-01-01
通讯作者: 王银叶*

Thrombolytic effect of rAcAP5 and its mechanism

Shuang Ding, Xiao-Yan Liu, Yuan-Jun Zhu, Yin-Ye Wang^*

Department of Molecular and Cellular Pharmacology, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China

Received:2011-05-09 Revised:2011-09-20 Online:2012-01-01 Published:2012-01-01
Contact: Yin-Ye Wang*

摘要/Abstract

摘要： 犬钩虫抗凝肽 (Ancylostoma caninum anticoagulant peptide, AcAP5) 是一种已报道的FXa抑制剂, 它对防止血栓溶解的TAFIa具有很强的抑制作用, 本文进一步研究rAcAP5的纤溶活性和溶栓活性。采用发色产物法测定rAcAP5对TAFIa的抑制作用, 用比浊法测定rAcAP5对尿激酶 (UK) 诱导的纤维蛋白溶解时间的影响, 通过血栓称重法进行体外溶栓实验和体内动静脉旁路溶栓实验。采用常规方法测定正常大鼠优球蛋白溶解时间 (ELT), 血浆纤维蛋白原含量和纤维蛋白降解产物 (FDP)含量。rAcAP5浓度依赖地抑制TAFIa活性, 其IC50为63.7 nmol/L, 为一个强的TAFIa抑制剂。rAcAP5 (5-40 nmol/L) 能够显著地加速尿激酶诱导纤维蛋白的溶解, 缩短纤溶时间。rAcAP5能够提高尿激酶诱导的血栓减重 (P<0.05), 单独使用rAcAP5也能显著增加体外实验的血栓减重 (P<0.05)。在大鼠体内的动静脉旁路模型中, 单次静脉给予rAcAP5 (50-200 μg/kg) 能够剂量依赖地增加血栓减重 (P<0.01)。rAcAP5在有效剂量对正常大鼠的ELT、血浆纤维蛋白原含量和FDP含量均无明显影响。研究结果提示: rAcAP5是一个强效溶栓多肽, 它可能通过抑制TAFIa的活性而在血栓表面具有溶栓活性, 而不影响循环血液中的纤溶活性和纤维蛋白原含量。

关键词: rAcAP5, TAFIa, 纤维蛋白溶解, 溶栓活性

Abstract: Ancylostoma caninum anticoagulant peptide 5 (AcAP5) has been reported as an FXa inhibitor. We found that this peptide showed potent inhibitory activity on activated thrombin-activatable fibrinolysis inhibitor (TAFIa), which protects the fibrin clot against lysis. This study investigated the effects of recombinant AcAP5 (rAcAP5) on fibrinolytic activity in vitro and on thrombolytic activity in vitro and in vivo. In addition, euglobulin lysis time (ELT), fibrinogen content and fibrin degradation product (FDP) in normal rat plasma were all determined to evaluate the influence of rAcAP5 on fibrinolytic activity in circulation blood. TAFIa activity was detected by colorimetry; fibrinolysis in vitro was determined with turbidimetry. Thrombolysis in vitro was measured by thrombus weight reduction; thrombolysis in vivo was conducted by arteriovenous shunt method with thrombus weight reduction. ELT, fibrinogen content and FDP were detected using routine method. rAcAP5 concentration-dependently inhibited TAFIa activity in vitro with an IC50 of 63.7 nmol/L. rAcAP5 (5-40 nmol/L) significantly accelerated urokinase-induced clot lysis of rabbit plasma and shortened fibrinolysis time (P<0.01). rAcAP5 at 1.5 μmol/L enhanced the reduced weight of thrombus induced by urokinase (P<0.05), and it also reduced thrombus weight (P<0.05) in vitro when used alone. In an arteriovenous shunt thrombolytic model, rAcAP5 (50-200 μg/kg, i.v.) dose-dependently enhanced the reduced weight of the thrombus. rAcAP5 at effective doses failed to influence ELT, fibrinogen contents and FDP in normal rat circulation plasma. In conclusion, rAcAP5 is a thrombolytic peptide, which may be attributed to its TAFIa inhibition. rAcAP5 may only exert thrombolytic activity on the thrombus site but not influence the fibrinolysis activity and fibrinogen in circulation plasma. These findings suggest that rAcAP5 is a potential thrombolytic candidate agent.

Key words: rAcAp5, TAFIa, Clot lysis, Thrombolytic activity

中图分类号:

R965

Supporting:

Foundation items: Original New Drug Fund of Peking University-Funder and National Technology Graveness Special Purpose Fund (Grant No. 2009zx09301-010).
^*Corresponding author. Tel.: 86-10-82802652; Fax: 86-10-62015584

丁双, 刘晓岩, 朱元军, 王银叶^*. rAcAP5的溶栓作用及作用机制研究[J]. , DOI: 10.5246/jcps.2012.01.009.

Shuang Ding, Xiao-Yan Liu, Yuan-Jun Zhu, Yin-Ye Wang^*. Thrombolytic effect of rAcAP5 and its mechanism[J]. , DOI: 10.5246/jcps.2012.01.009.

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