Preparation and antithrombotic activity identification of Perinereis aibuhitensis extract: a high temperature and wide pH range stable biological agent.
Preparation and antithrombotic activity identification of Perinereis aibuhitensis extract: a high temperature and wide pH range stable biological agent.
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DOI：
10.1039/c7fo00987a
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发表时间：
2017-10
期刊：
影响因子：
6.1
通讯作者：
Yang Li;Jing Li;Tianhong Liu;Ying Wang;Zhongzheng Zhou;Feng Cheng;Chao Feng;Xiaojie Cheng
中科院分区：
文献类型：
--
作者：
Yang Li;Jing Li;Tianhong Liu;Ying Wang;Zhongzheng Zhou;Feng Cheng;Chao Feng;Xiaojie Cheng
本工作从沙蚕中提取了一种热稳定性好、pH范围宽(1~13)的生物制剂，并对其抗血栓活性进行了研究。体外抗血栓活性实验表明，Pae能显著延长兔血复钙后的凝血时间：Pae 10、20、40 mg m L-1组的凝血时间分别增加24.24%、68.64%和130.76%。进一步的血小板贫乏血浆(PPP)凝血实验证明，PAE对内源性凝血途径有明显的抑制作用，而对外源性凝血途径的抑制作用较弱。此外，Pae还能有效抑制凝血酶诱导的纤维蛋白原转化为纤维蛋白。更重要的是，由于PAE在制备过程中被煮熟，因此PAE具有热稳定性，并且由于其在广泛的pH值条件下处理后保持了对纤维蛋白原的抑制活性，因此具有较强的耐酸性和耐碱性。最后，对PAE中的抗血栓活性成分进行了初步鉴定，结果表明该成分不溶于乙醇，对木瓜酶和胃酶不敏感，可被H_2O_2降解。此外，通过体外溶血率试验和四甲基偶氮唑盐比色试验证明，PAE在不超过20mgmL-1的浓度下表现出较高的抗凝血活性，具有良好的生物相容性。综上所述，该PAE制备方便、稳定性好，是一种安全有效的抗血栓药物，具有开发抗血栓食品的潜力。
In this work, a thermal and wide pH range (from 1 to 13) stable biological agent was extracted from Perinereis aibuhitensis (P. aibuhitensis), and the antithrombotic activity of this P. aibuhitensis extract (PaE) was investigated. In vitro antithrombotic activity test indicated that PaE could significantly extend the clotting time of recalcified rabbit blood: the clotting time of the 10 mg mL-1, 20 mg mL-1 and 40 mg mL-1 PaE groups increased by 24.24%, 68.64% and 130.76%, respectively. Further platelet poor plasma (PPP) coagulation study proved that PaE could obviously inhibit the intrinsic coagulation pathway and slightly inhibit the extrinsic coagulation pathway. Moreover, PaE could effectively inhibit fibrinogen turning into fibrin induced by thrombin. More importantly, PaE exhibited thermal stability, since P. aibuhitensis was boiled in the PaE preparation process, as well as tolerance to strong acid and alkali, since its fibrinogen inhibition activity was retained after treatment with a wide range of pH value conditions. Finally, the antithrombotic activity ingredients in PaE were preliminarily identified, the results of which indicated that the ingredients were ethanol insoluble, insensitive to papain and pepsin, and could be degraded by H2O2. Additionally, we demonstrated that PaE at a concentration of not more than 20 mg mL-1, the concentration at which PaE showed high anticoagulant activity, exhibited favorable biocompatibility via in vitro hemolysis rate test and MTT assay. In summary, the results evidenced that this conveniently prepared and stable PaE was a safe and effective antithrombotic agent, and had the potential to be utilized for antithrombotic food development.