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

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

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(K_b),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,K_b为3.39×104L/mol,FA与B-Thr的K_b为1.05×104L/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K_b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。

Investigation on interactions between bovine thrombin （B-Thr）, aptamer29 （Apt29） and three active components （salvianic acid A sodium （SAS）,protocatechuic acid （PA）, feru-le acid （FA）） in Danshen was carried out by capillary zone electrophoresis （CZE）. CZE condi-tions 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 （瓦b） 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 Kh 3. 39× 10^4 L/mol. Kh between FA and B-Thr was 1. 05×10^4 L/mol. SAS had no binding with B- Thr. FA showed stron-ger affinity with Apt29 than PA with Kh of 1. 48×10^4 L/mol and 1. 32×10^4 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 （pro-tein） and herbal drug molecules. And it could give insight for target discovery and delivery of herbal drug molecules.