药学学报, 2019, 54(12): 2178-2183
引用本文:
王玄, 甘奇超, 史景彦, 胡黄婉茵, 郝俊杰, 邬国松, 高倩, 李响, 牛明, 王伽伯, 马莉, 肖小河. 水蛭抗血小板聚集生物效价检测方法研究[J]. 药学学报, 2019, 54(12): 2178-2183.
WANG Xuan, GAN Qi-chao, SHI Jing-yan, HU Huang-wan-yin, HAO Jun-jie, WU Guo-song, GAO Qian, LI Xiang, NIU Ming, WANG Jia-bo, MA Li, XIAO Xiao-he. Measurement of antiplatelet aggregation and potency of hirudo[J]. Acta Pharmaceutica Sinica, 2019, 54(12): 2178-2183.

水蛭抗血小板聚集生物效价检测方法研究
王玄1,2, 甘奇超3, 史景彦3, 胡黄婉茵2, 郝俊杰2, 邬国松2, 高倩1, 李响1, 牛明2, 王伽伯2, 马莉1, 肖小河2
1. 首都医科大学中医药学院, 北京 100069;
2. 解放军总医院第五医学中心, 全军中医药研究所, 中西医结合诊疗与研究中心, 北京 100039;
3. 重庆多普泰制药股份有限公司, 重庆 401420
摘要:
建立基于抗血小板聚集活性的水蛭生物效价测定法,用于补充其质量控制方法。凝血酶滴定法测定高温煎煮后水蛭的抗凝血酶活性,对凝血酶定法进行适用性再评价。测定水蛭体内和体外的抗血小板聚集活性。以血小板聚集抑制率为指标,优化水蛭提取方法,建立新的生物效价测定法。进行方法学考察,测定12批水蛭样品的效价。实验过程符合动物实验伦理标准。结果表明水蛭经高温煎煮后抗凝血酶活性显著降低,几乎丧失活性。药效实验表明水蛭具有抗血小板聚集活性,体内外结果一致。方法学考察结果均符合要求。12批样品的效价(PT)分别为113.494、96.126、121.224、127.326、83.482、108.72、131.409、127.946、76.899、126.266、132.894、573.525 U·mg-1。结果提示基于抑制血小板聚集活性的水蛭生物效价测定法方法可靠,重复性良好,可用于其质量评价。
关键词:    水蛭      活血化瘀      抗血小板聚集      生物效价      质量评价     
Measurement of antiplatelet aggregation and potency of hirudo
WANG Xuan1,2, GAN Qi-chao3, SHI Jing-yan3, HU Huang-wan-yin2, HAO Jun-jie2, WU Guo-song2, GAO Qian1, LI Xiang1, NIU Ming2, WANG Jia-bo2, MA Li1, XIAO Xiao-he2
1. School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China;
2. China Military Institute of Chinese Medicine, Integrative Medical Center, Fifth Medical Center, General Hospital of Chinese PLA, Beijing 100039, China;
3. Chongqing Duoputai Pharmaceutical Co., Ltd., Chongqing 401420, China
Abstract:
The objective of this work was to develop a bioassay to quantify the antiplatelet aggregation activity of hirudo for quality evaluation and control. Antithrombin activity of hirudo extracted by high temperature decoction was determined by thrombin titration. Antiplatelet aggregation activity of hirudo was determined through pharmacodynamic experiments in vitro and in vivo using a bioassay we developed for quantifying inhibition of platelet aggregation. Methodological investigation was carried out and the titers of 12 batches of hirudo samples were determined. During the experiment, the disposal of animals is in accordance with the ethical standards of animal experiments. The results showed that the antithrombin activity of hirudo decocted at high temperature decreased significantly and almost lost its activity. Hirudo inhibited platelet aggregation and results in vivo and in vitro were consistent. These assays were employed to test 12 batches of hirudo. The results demonstrated that the biopotency of 12 batches was 113.49, 96.13, 121.22, 127.33, 83.48, 108.72, 131.41, 127.95, 76.90, 126.27, 132.89 and 573.53 U·mg-1. The method was reliable and reproducible and can be used to assess the quality of hirudo.
Key words:    hirudo    promoting blood circulation    anti-platelet aggregation    potency    quality evaluation   
收稿日期: 2019-03-02
DOI: 10.16438/j.0513-4870.2019-0147
基金项目: 国家自然科学基金资助项目(81673596);国家中药标准化项目(ZYBZH-C-HUN-21,ZYBZH-Y-TJ-43,ZYBZH-Y-BJ-07).
通讯作者: 牛明,Tel:86-10-66933325,E-mail:nmbright@163.com;王伽伯,Tel:86-10-66933325,E-mail:wjb0128@126.com;马莉,Tel:86-10-83911635,E-mail:marytcm@ccmu.edu.cn
Email: nmbright@163.com;wjb0128@126.com;marytcm@ccmu.edu.cn
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