基于P糖蛋白核酸适配体分子探针的傣药龙血竭脑靶向转运及抗脑缺血药效物质基础研究

以血脑屏障上主要外排蛋白P糖蛋白（P-gp）为靶标，筛选其特异性抑制剂-核酸适配体AptP-gp。结合微透析采样及液质联用技术，开展傣药龙血竭、AptP-gp、P-gp三者间体内、体外相互作用研究。依据龙血竭与P-gp、AptP-gp间结合常数，阐明与AptP-gp特异结合的化学成分群、分析AptP-gp对龙血竭多成分脑靶向运输能力。将AptP-gp作为竞争分子探针，识别哪些成分为P-gp潜在底物、确定P-gp是否为龙血竭进入鼠脑的主要外排蛋白。利用AptP-gp对血脑屏障P-gp精准定位和促龙血竭跨血脑屏障转运能力，阐明龙血竭抗脑缺血药效物质基础。利用荧光标记的AptP-gp可视化识别龙血竭鼠脑内靶点定位。本项目将新兴热点功能分子-核酸适配体引入到傣药研究中，基于其独特性能研究傣药多成分脑靶向运输、靶点定位、效应成分群识别，具有明显创新性，或为傣药基础研究提供可借鉴的新思路及新方法。

Specific aptamers (AptP-gp) are selected using P-glycoprotein (P-gp) as target, which is the main efflux transporter in brain-blood barrier. Study on in vitro and in vivo interaction between Dragon’s blood, AptP-gp and P-gp will be performed with microdialysis and HPLC-MS methods. The chemical constituents specifically binding AptP-gp and P-gp，the potential substrates of P-gp could be elucidated by determining combination constants between AptP-gp/P-gp and multi-components of Dragon’s blood. The AptP-gp brain targeting ability could be evaluated by changed levels of multi-components of Dragon's Blood in rat brain with addition of AptP-gp. Whether P-gp is the efflux transporter for multi-components will be determined with AptP-gp as competitive binding probe. Anti-ischemic material basis will be elucidated by precise localization and targeting delivery capacities of AptP-gp. Targeting sites of Dragon’s blood in rat brain could be analyzed by fluorescence-labeled AptP-gp. By introducing the newly emerged functional molecule-aptamer into basic research of Dai medicine, study on multi-component targeting delivery, localization and therapeutic material basis identification using aptamers prove to be significantly innovative, which could provide a new idea and approach for traditional Chinese medicine research.