基于与核酸适配体相互作用研究龙血竭抗凝血靶点及物质基础

基于目前热点识别分子-核酸适配体的靶点定位及选择性运输功能，结合微透析采样及液质联用技术，首次开展龙血竭与人凝血酶抑制剂核酸适配体Apt 15、Apt 29及凝血酶间体内外相互作用研究。测定龙血竭多成分与凝血酶、Apt 15、Apt 29间结合常数，确定抑制凝血酶的化学成分群、与Apt 15、Apt 29特异结合的化学成分群。将Apt 15、Apt 29作为凝血酶位点竞争结合探针，确定龙血竭多成分是否作用于凝血酶肝素或纤维蛋白原结合位点。观察Apt 15、Apt 29对龙血竭多成分在鼠脑中含量变化的影响，结合药效，分析Apt 15、Apt 29对龙血竭多成分向鼠脑选择性运输能力和药效协同作用。本研究交叉运用传统中医药理论与现代分子生物学技术，基于核酸适配体研究中药多成分靶点定位、选择性运输、效应成分群识别，具有明显的创新性，或为中药相关基础研究提供可借鉴的新思路及新方法。

Based on the target binding and delivery functions of aptamer, the newly emerging recognition molecule, study on in vitro and in vivo interaction between Dragon's blood, thrombin inhibitor aptamers Apt 15 and Apt 29 and thrombin will be performed with microdialysis and HPLC-MS method. The chemical constituents with thrombin inhibitory effect will be elucidated by binding constants between thrombin and multi-components of Dragon's blood. Whether multi-components could bind heparin and fibrinogen exosites will be determined with Apt 15 and Apt 29 as competitive binding probes. The chemical constituents in Dragon's blood which could specifically bind with Apt 15 and Apt 29 will be elucidated. Effect on level of multi-components of Dragon's Blood in rat brain will be studied with the addition of Apt 15 and Apt 29. Target delivery to rat brain and synergism between aptamers and Dragon's blood will be observed with the pharmacological results. By cross-use of TCM theory and modern molecular biology techniques, study on multi-component target positioning ,selevtive delivery and material basis identification based on aptamers will provide a new idea and approach for TCM research with a significant innovation.