基于 "发汗解表、能量代谢、免疫调节"三位一体的"辛凉解表"方剂药效物质基础与作用机理研究

With the increasing weather haze and pollution, the incidence of respiratory diseases rises sharply in recent years. "Wenxie Fanfei" is the main pathogenesis. Meanwhile, modern medical in prevention and treatment of lung inflammation has encountered a bottleneck caused by increasing of drug-resistant bacteria. "Xinliang Jiebiao" is one of the common methods in traditional Chinese medicine in the treatment of "Wenxie Fanfei". Combined with Chinese medicine theory and modern science to interpret its connotation, might be an effective way to solve this problem. In our previous study, we found that there were some medicinal composition in Xinliang Jiebiao drugs could regulate the signaling pathways of ADR, ATP synthesis and NF-κB. All these effects were closely related with "sweating, energy metabolism and immune regulation". In this present study, systems biology research on Sang Ju Yin and Yin Qiao San were build a syndrome model of "Wenxie Fanfei". We try to identify the effective compounds rapidly by bioactivity-based UPLC-Q/TOF screening system; reveal its associated targets, pathways and networks using cell imaging and protein target fishing technology. Through the study of molecular mechanism of enhance the signal transduction by the prescription compatibility, the scientific connotation of compound compatibility synergies will be investigated, and the scientific justification for adapting the remedy to "Wenxie Fanfei" by traditional Chinese medicine might be clarified.

呼吸系统疾病的发病率近年来随着大气污染、雾霾天气的增加而急剧上升，"温邪犯肺"是其主要病机之一，同时现代医学对其防治因耐药菌增多等因素而遭遇了瓶颈。"辛凉解表"是中医治疗温邪犯肺的常用治法，将中医理论与现代科学相结合诠释其内涵，将为温热型肺病的治疗提供科学依据及有效途径。我们前期研究发现辛凉解表药物中存在调控ADR、ATP synthesis、NF-κB等关键信号通路的药效物质，其药理作用与"发汗解表、能量代谢、免疫调节"密切相关。本项目通过构建温邪犯肺证候模型，开展辛凉解表代表方桑菊饮、银翘散的系统生物学研究；结合活性导向的UPLC-Q/TOF快速筛选平台，辨析辛凉解表药的药效成分；以化学生物学手段标记的关键成分进行示踪及靶点垂钓，揭示其作用靶点、通路以及网络的关联；通过方剂配伍增强相关信号转导的分子机制研究，阐述经典方剂配伍协同增效的科学内涵，阐释中医治疗"温邪表证"对证施药的科学性。

随着气候的变化以及雾霾和污染天气的增加，呼吸系统疾病发病率急剧上升，而现代医学对耐药细菌以及病毒感染等引起的肺部炎症的防治又遇到了瓶颈。“辛凉解表”是中医治疗温邪犯肺的常用治法，如何将中医理论与现代科学相结合，诠释其内涵是解决当前面临疾患的有效途径之一。.我们在前期研究基础之上预测辛凉解表之“发汗解表、能量代谢、免疫调节”作用的科学内涵与NF-κB、Ca2+、ADR、能量代谢等多种途径调控相关。在本项目中，我们以从系统生物学到化学生物学的研究理念为指导，首先对银翘散、桑菊饮等辛凉解表经典方药整体生物活性进行评价，并进一步利用前期建立的生物活性导向UPLC-Q/TOF快速筛选平台对经典复方及单味解表药中相关活性成分进行筛选，结果显示经典方药中含有较多NF-κB调控成分（连翘酯苷E、绿原酸、隐绿原酸，马钱苷等）、部分具有解表、清热作用的中药中存在调控ADR（麻黄碱、升麻酰胺等）及Ca2+（连翘苷元、甘草次酸等）成分，另外也有部分辛凉解表中药中存在调控ATP成分（连翘苷元、甘草次酸）。最后选择连翘苷元、绿原酸、甘草次酸等关键药效成分，对其进行结构修饰，应用化学生物学方法捕获其直接作用靶点，采用蛋白组学方法对其靶点进行鉴定，并结合多种分子生物学手段进行功能确证。结果显示，连翘苷元、绿原酸等化合物抗炎作用与直接调控AKT、Ras蛋白密切相关；连翘苷元、甘草次酸等化合物钙拮抗作用与直接调控PLC、钙通道蛋白密切相关。本研究结果提示辛凉解表方剂作用机制与多成分、多靶点的调控NF-κB、Ca2+、ADR、能量代谢，进而发挥“发汗解表、能量代谢、免疫调节”作用相关。本研究为阐述辛凉解表经典方药作用机制的科学内涵，进而证明中医治疗“温邪表证”对证施药的科学性提供了实验依据。.在本项目资助下，现已发表论文13篇，其中SCI收录10篇；培养博士毕业生2名，硕士毕业生5名。达到了预期研究目标。