色谱 ›› 2017, Vol. 35 ›› Issue (3): 339-343.DOI: 10.3724/SP.J.1123.2016.10018

• 研究论文 • 上一篇    下一篇

毛细管区带电泳法考察牛凝血酶、核酸适配体与丹参活性分子间的相互作用

李玉娟, 武广霞, 欧婉露, 孙欣欣, 屈锋   

  1. 北京理工大学生命学院, 北京 100081
  • 收稿日期:2016-10-09 出版日期:2017-03-08 发布日期:2013-07-16
  • 通讯作者: 李玉娟,E-mail:lylyjlzh2006@163.com;屈锋,E-mail:qufengqu@bit.edu.cn.
  • 基金资助:

    国家自然科学基金项目(81573693,21675012,21375008).

Investigation on interactions between bovine thrombin, aptamer and active components in Danshen by capillary zone electrophoresis

LI Yujuan, WU Guangxia, OU Wanlu, SUN Xinxin, QU Feng   

  1. School of Life Science, Beijing Institute of Technology, Beijing 100081, China
  • Received:2016-10-09 Online:2017-03-08 Published:2013-07-16
  • Supported by:

    National Natural Science Foundation of China (Nos. 81573693, 21675012, 21375008).

摘要:

采用毛细管区带电泳法(CZE)研究牛凝血酶(B-Thr)、凝血酶核酸适配体(Apt29)与丹参中3种活性物质(丹参素钠(SAS)、原儿茶酸(PA)、阿魏酸(FA))间的相互作用。电泳条件为:毛细管长度40 cm,紫外检测波长214 nm,分离电压15 kV,压力进样3.45 kPa,进样时间5 s。分别固定3种活性药物分子的浓度,改变B-Thr或Apt29的浓度,采用Scatchard方程和非特异性结合方程计算结合常数(Kb),表征药物分子和B-Thr或Apt29的相互作用大小。结果表明,PA与B-Thr结合能力最强,Kb为3.39×104L/mol,FA与B-Thr的Kb为1.05×104 L/mol,SAS与B-Thr未表现出结合能力;在与Apt29的相互作用的研究中显示,SAS基本未与Apt29结合,FA与Apt29结合能力强于PA与Apt29的结合能力,FA、PA与Apt29的Kb分别为1.48×104L/mol和1.32×104L/mol。该研究报道了丹参活性分子与B-Thr、Apt29的相互作用结果,可为核酸适配体(蛋白质)与中药分子相互作用的探索提供新方法,适配体与中药分子相互作用亦可为中药分子的靶点发现、靶向转运提供借鉴。

关键词: 丹参, 核酸适配体, 毛细管区带电泳法, 牛凝血酶

Abstract:

Investigation on interactions between bovine thrombin (B-Thr), aptamer29 (Apt29) and three active components (salvianic acid A sodium (SAS), protocatechuic acid (PA), ferulic acid (FA)) in Danshen was carried out by capillary zone electrophoresis (CZE). CZE conditions were as follows: capillary length of 40 cm, voltage of 15 kV, injection volume of 3.45 kPa, injection time of 5 s. By changing concentration of B-Thr or Apt29 with fixed levels of the three active components, binding constants (Kb) were calculated by Scatchard and non-specific binding equations to show interactions between B-Thr, Apt29 and the three active components. The results showed that PA had the strongest affinity to B-Thr with Kb 3.39×104L/mol. Kb between FA and B-Thr was 1.05×104 L/mol. SAS had no binding with B-Thr. FA showed stronger affinity with Apt29 than PA with Kb of 1.48×104 L/mol and 1.32×104 L/mol, respectively. The present study reported that interactions between the active constituents in Danshen and B-Thr/Apt29. New methods might be supplied for calculation of interactions between Apt (protein) and herbal drug molecules. And it could give insight for target discovery and delivery of herbal drug molecules.

Key words: aptamer (Apt29), bovine thrombin (B-Thr), capillary zone electrophoresis (CZE), Danshen

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