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

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (12): 1355-1360.

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

黄璐¹, 张根葆^1,2, 闵志雪¹, 马开然¹, 郑汝琦¹, 孙瑶¹, 吴娟¹

¹皖南医学院病理生理学教研室,²蛇毒研究所,芜湖 241002,安徽

收稿日期:2012-09-03 修回日期:2012-11-05 出版日期:2012-12-26 发布日期:2012-12-31
作者简介:黄璐,女,硕士研究生,研究方向:心血管病理生理学。Tel: 18255354275 E-mail: xidianlulu1986@qq.com
基金资助:
安徽省教育厅自然科学基金重点资助项目(KJ2011A266)

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

HUANG Lu¹, ZHANG Gen-bao^{1, 2}, MIN Zhi-xue¹, MA Kai-ran¹, ZHENG Ru-qi¹, SUN Yao¹, WU Juan¹

¹Department of Pathophysiology; ²Institute of snake venom, Wannan Medical College, Wuhu 241002, Anhui, China

Received:2012-09-03 Revised:2012-11-05 Online:2012-12-26 Published:2012-12-31

摘要/Abstract

摘要： 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,5 mg/kg),AHV-PI(0.1 mg/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。

关键词: 蝮蛇毒, 血小板抑制因子, 动脉血栓, 血小板

Abstract: 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 (P<0.01 and P<0.05). The levels of GMP-140 and TXB₂ in plasma were decreased (P<0.01). 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.

Key words: Agkistrodon halys venom, Platelet inhibitor, Artery thrombosis, Platelet

中图分类号:

R965.2

黄璐, 张根葆, 闵志雪, 马开然, 郑汝琦, 孙瑶, 吴娟. 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[J]. 中国临床药理学与治疗学, 2012, 17(12): 1355-1360.

HUANG Lu, ZHANG Gen-bao, MIN Zhi-xue, MA Kai-ran, ZHENG Ru-qi, SUN Yao, WU Juan. Effect and mechanism of platelet inhibitor from Agkistrodon halys venom on artery thrombosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(12): 1355-1360.

参考文献

[1] 王振义, 李家增, 阮长耿, 等. 主编. 血栓与止血-基础理论与临床[M].上海: 上海科学技术出版社, 2004:460-464.
[2] Ouyang C, Teng CM, Huang TF.Characterization of snake venom components acting on blood coagulation and platelet function[J]. Toxicon, 1992, 30(9): 945-966.
[3] 冯敏, 赵洪洋, 刘莹莹, 等. 家兔实验性血栓模型的建立及其检测[J]. 临床心血管杂志, 2009, 25(10): 781-783.
[4] Anderews PK, Shen Y, Gardiner EE.The glycoprotein Ib/IX/V complex in platelet adhesion and signaling[J]. Thromb Haemost, 1999, 82(3): 357-364.
[5] Matsui H, Sugimoto M, Mizuno T, et al.Distinct and concerted functions of von Willebrand facter and fibrinogen in mural thrombus growth under high shear flow[J]. Blood, 2002, 100(24): 3604-3610.
[6] 庞兴学, 王显. 血栓形成的过程与机制研究进展[J]. 医学综述, 2011, 17(11): 613-616.
[7] Ouyang C, Huang TF.Inhibition of platelet aggregation by 5,-nucleotidase purified from Trimeresurus gramineus snake venom[J]. Toxicon,1983,21(4):491-498.
[8] 袁帅, 刘淑清, 付冬雪, 等. 长白山白眉蝮蛇蛇毒酸性PLA2质谱鉴定及其对血小板聚集的抑制作用[J]. 生物学杂志, 2010, 27(2): 13-15.
[9] 班建东, 蓝秀万, 舒雨雁. 蝮蛇毒中纤溶组分Ⅱ对血小板聚集的影响[J]. 临床荟萃, 2001, 16(9): 391-392.
[10] Storey RF, May JA, Wilcox RG, et al.A whole blood assay of inhibition of platelet aggregation by glycoprotein IIb/IIIa antagonists:comparison with other aggregation methodologies[J]. Thromb Haemost, 1999, 82(4): 1307-1311.
[11] 段婷, 张根葆, 周兵. 皖南蝮蛇毒蛋白C激活物的急性毒性作用的研究[J]. 中国临床药理学与治疗学, 2012, 17(1): 20-24.
[12] Michelson AD, Barnard MR, Krueger LA, et al.Circulating monocyte-platelet aggregates are a more sensitive marker of in vivo platele activation than platelet surface P-selectin: studies inbaboons, human coronary intervention, and human acute myocardial infarction[J]. Circulation, 2001, 104(13): 1533-1537.
[13] Palikhe NS, Kim SH, Lee HY, et al.Association of thromboxaneA2 receptor (TBXA2R) genepolymorphism in patients with aspirinintolerant acute urticaria[J]. Clin ExpAllergy, 2011, 41(2): 179-185.
[14] 钟灼, 观美华, 陈爱, 等. 奥扎格雷钠对高凝状态者凝血和纤溶指标的影响[J]. 临床血液学杂志, 2001, 14(6): 249-252.

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