融合体内外分析—靶标网络的藏药“达布班杂”治疗HAPE的药效物质及作用机制研究

High Altitude Pulmonary Edema (HAPE) is a commonly and frequently occurring acute mountain sickness, which is a serious threat to people’s health in plateau area. The previous study (No. 81503361) of our research group showed that the main medicine of “Da Bu Ban Za” (Wuwei Shaji San)-Shajigao has preventive and therapeutic effects of HAPE. Therefore, this project proposed a hypothesis which is “the effective substances and mechanisms of Tibetan medicine ‘Da Bu Ban Za’ in the treatment of HAPE, can be accurately identified by the combined research strategy of in vivo-vitro bioactive compounds analysis and target network” based on the theory and experience of Tibetan medicine. Firstly, the effective substances will be studied by the comparative analysis of in vivo and vitro UPLC-Q-TOF/MS chemical fingerprints. The network model of effective substances, targets and HAPE will be subsequently established and studied using network pharmacology. At the same time, the network model will be verified by the expression of ROS/HIF-1α/VEGF-A in hypoxia induced HPMEC injury model. The effective substances and mechanisms of “Da Bu Ban Za” in the treatment of HAPE will be deeply explained on the basis of the aforementioned results, and the scientific content of classic Tibetan medicine will be comprehensively disclosed.

高原肺水肿（HAPE）是一种急发性重症高原病，严重威胁进入高原地区人民的身体健康。本课题组前期研究（No. 81503361）发现，藏药达布班杂（五味沙棘散）方中主药—沙棘膏对HAPE具有较好的防治作用。故本项目结合藏医药理论和用药经验，提出“融合体内外药效成分分析—靶标网络的研究策略，能够准确识别达布班杂治疗HAPE的药效物质与作用机制”的假说。通过体外药物、高原低氧环境诱导HAPE动物模型血清的UPLC-Q-TOF/MS化学指纹图谱比对分析，筛选其药效物质。并以此作为切入点，通过网络药理学方法寻找其作用靶点，构建并分析达布班杂药效物质与HAPE靶标相互作用的网络模型。同时，结合达布班杂对低氧诱导HPMEC损伤模型ROS/HIF-1α/VEGF-A表达的影响进行验证，以深入诠释藏药经典复方达布班杂治疗HAPE的药效物质与作用机制，揭示其多成分、多靶点、多途径作用的科学内涵。

本课题针对藏药多成分、多靶点、多途径的作用特点，延续前期研究，以藏药经典复方达布班杂为研究对象，首先对方中多基原藏药沙棘进行基于Bar-HRM的分子生物学鉴定及基于tri-step IR的化学鉴定研究，实现从基原植物、药材及混伪品等用药形式的准确鉴定；通过对达布班杂的体外药物、高原低氧环境诱导HAPE动物模型血清的UPLC-Q-TOF/MS化学指纹图谱比对分析，筛选指认槲皮素、山奈素、异鼠李素、没食子酸、鞣花酸、柯里拉京、木香烃内酯、去氢木香内酯、甘草酸、甘草次酸、甘草苷、甘草素、异甘草素、紫檀芪等为该复方治疗HAPE的主要药效物质；以此作为切入点，通过网络药理学方法分析确认达布班杂治疗HAPE的26个核心靶点，主要涉及蛋白、酶、核酸结合、受体、转录因子、信号分子、酶调节器等，构建的“化合物-核心靶点-通路”网络模型包含30个关键活性成分，作用于20个核心靶点，31条关键通路，主要涉及炎症反应、免疫系统、氧化应激等多种途径。达布班杂对低氧诱导HPMECs损伤模型ROS/HIF-1α/VEGF-A表达影响的验证实验表明，该复方能够降低发生炎性损伤HPMVECs中ROS的含量，以及HIF-1α和VEGF-A蛋白表达，提示该藏药复方可为预防和治疗HAPE发挥积极的作用，诠释了藏药经典复方达布班杂治疗HAPE的药效物质与作用机制，揭示其多成分、多靶点、多途径作用的科学内涵，也为其治疗HAPE在临床上的推广应用和进一步开发提供可借鉴的科学依据。本项目严格按照预期计划开展研究，取得了较好的研究成果，共发表学术论文4篇，其中SCI论文2篇，中文核心期刊论文2篇；相关研究成果获得2019年度中国民族医药学会科学技术奖一等奖1项。