基于多元核酸适配体生物探针研究傣药龙血竭抗失重肠上皮屏障损伤的靶点网络与体内药效物质

龙血竭是傣医用于治疗机体水塔病变的传统傣药，课题组多年研究发现其能有效对抗航天失重导致的肠上皮屏障损伤，但药效物质及作用机制尚不明确，严重制约了其在航天医学领域的应用。本项目以肠上皮连接中闭锁蛋白（OCN）、闭锁小带蛋白（ZO-1）为靶标，筛选各自核酸适配体APT，作为多元生物探针。利用APT具有靶点定位、与靶标特异性结合、在体内与龙血竭选择性竞争靶标的功能，结合磁分离、液质联用及差异蛋白组学技术，可特异性阐明与OCN\ZO-1结合的龙血竭成分群、精准揭示调控模拟失重鼠肠上皮连接损伤的靶点网络及作用机制、靶向性发现龙血竭作用在肠上皮连接OCN\ZO-1信号网络的物质基础。本项目将新兴功能分子核酸适配体、现代分子生物学技术、传统医药理论有机结合，从分子水平研究傣药龙血竭保护模拟失重鼠肠上皮屏障损伤的靶点网络和体内药效成分群，具有明显创新性，亦或为傣药基础研究提供可借鉴的新方法。

Dragon’s Blood (DB) is one kind of traditional Dai Medicine and Dai doctors use it to treat “Water Tower” diseases. Many years’ research in our group has shown that DB could significantly prevent intestine epithelial barrier injury induced by microgravity. However, therapeutic materials and action mechanism remain unclear, which seriously restricts its application in the field of space medicine. Using Occludin (OCN) and Zonula Occluden-1 (ZO-1) in intestine epithelial barrier as targets, APT(OCN) and APT(ZO-1) are selected as multi biological probes. Using the ability of APT target localization, specifically binding with targets and competitively binding targets with DB in vivo, chemical components in DB binding with OCN\ZO-1 would be specifically identified. Target network and mechanism of DB acting on intestine epithelial junction in simulated microgravity rats could be precisely elucidated. Material basis of DB acting on OCN\ZO-1 signaling network in rat intestine epithelial junction would be specifically screened with magnetic separation, HPLC-MS technique and differential proteomics. By cross-use of new-emerged APT functional molecules, modern molecular biology techniques and theory of traditional medicine, study on target network and in vivo therapeutic constituents of DB preventing rat intestine epithelial barrier injury induced by simulated microgravity would be investigated at the molecular level, which proves to be significantly innovative, and might provide new method for fundamental research of Dai Medicine.