Anticoagulant activity of crude extract of Holotrichia diomphalia larvae
Anticoagulant activity of crude extract of Holotrichia diomphalia larvae
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大黑鳃金龟幼虫粗提物的抗凝血活性
DOI：
10.1016/j.jep.2015.11.015
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发表时间：
2016-01-11
影响因子：
5.4
通讯作者：
Liu, Shuwen
中科院分区：
文献类型：
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作者：
Xu, Xueqing;Liu, Wenjun;Liu, Shuwen
民族药理学相关性：东北大黑鳃金龟（Holotrichia diomphalia larvae）是东亚民间的一种经典药材，临床上用于活血化瘀已有数百年的历史。材料与方法：采用正常人血浆、富浆血小板、鼠尾离体和牛纤维蛋白原，测定了CEHDL对血浆凝固时间、血小板聚集、出血时间、纤维蛋白原和纤维蛋白原水解的影响;采用血栓溶解实验和角叉菜胶诱导小鼠尾血栓形成模型，研究CEHDL的体内外抗血栓形成活性。CEHDL含有大量的蛋白质，能抑制血液凝固和血小板聚集，并呈剂量依赖性。此外，CEHDL优先切割纤维蛋白原的α链和β链，然后切割γ链。此外，CEHDL可直接降解纤维蛋白，而不激活纤溶酶原。已经注意到CEHDL的纤溶活性可以不受金属离子如Mg 2+、Ca 2+、Zn 2+、Fe 2+、Fe 3+、Cu 2+和pH 3 - 10的缓冲液的影响。蛋白酶抑制剂如TPSI、抑肽酶、亮皮素、PMSF、DTT和EDTA对CEHDL的纤溶活性只有轻微的抑制作用或无抑制作用，但在75 ℃和100 ℃下CEHDL可完全失活。结论：CEHDL具有较强的抗凝血活性，并具有多种血纤维蛋白原溶解活性。其抗血栓作用与其作为药物的作用有关。(c)2015爱思唯尔爱尔兰有限公司版权所有。
Ethnopharmacological relevance: Holotrichia diomphalia larvae are one classical folk medicinal material in East Asia which has clinically been used to promote blood circulation and dispel blood stasis for several hundred years.Aim of the study: The anticoagulant activity of crude extract of H. diomphalia larvae (CEHDL) in vitro and in vivo was evaluated to explore its mechanism as antithrombotic medicine.Materials and methods: The effects of CEHDL on plasma recalcification time, platelet aggregation, bleeding time, hydrolysis of fibrinogen and fibrin were measured with normal human plasma, plasmarich platelet, transected mouse tails and bovine fibrinogen; the anti-thrombosis activities of CEHDL in vitro and in vivo were analyzed with clots lysis assay and carrageenan-induced mouse tail thrombosis model.Results: CEHDL was found to contain large numbers of proteins and could inhibit blood coagulation and platelet aggregation in a dose-dependent manner. Furthermore, CEHDL preferentially cleaved alpha- and beta-chains followed by gamma-chains of fibrinogen. Besides, CEHDL could directly degrade fibrin rather than activate plasminogen. It has been noted that fibrinogenolytic activity of CEHDL could be unaffected by metal ions such as Mg2+, Ca2+, Zn2+, Fe2+, Fe3+, Cu2+ and buffers with pH 3-10. Moreover, protease inhibitors like TPSI, aprotinin, leupetin, PMSF, DTT and EDTA only slightly or not inhibited fibrinogenolytic activity of CEHDL However, CEHDL could be completely inactivated at 75 degrees C and 100 degrees C. In addition, CEHDL exhibited anti-thrombosis activities in both blood clot lysis assay and carrageenan-induced thrombosis model.Conclusion: CEHDL possessed potent anticoagulant activity and several fibrin(ogen)olytic agents from H. diomphalia larvae were responsible for its antithrombotic effect as medicine. (c) 2015 Elsevier Ireland Ltd. All rights reserved.