基于光偶联探针研究栀子成分肝保护及肠道菌药效机制

Confirmation of biologically active molecules of Chinese medicine (CM) in treating or regulating human diseases plays a crucial role in in-depth investigation of CM. However, active chemicals in CM, as well as target CM gets involved in, have still remained unknown. Therefore, this state drastically undermine modernization and internationalization of CM. Our previous pharmacological studies revealed that gardenia possess significant properties of hepatic protection and regulation of intestinal microbiota. Therefore, in current project, we aim to design and synthesize plotoactive probes by means of chemical biology, and then combine them with main metabolites of gardenia. Then, obtained compounds would be incubated with target organ cells and intestinal bacteria, and lysate would be treated by UV irradiation. After that, proteomics of extracted proteins by gel separation would be analyzed comprehensively to uncover compounds which react with target proteins from hepatic cells and intestinal bacteria, and determine target proteins. Generally, the protocol of current project is not simply based upon addition of various disciplines, but highly integration of a variety of disciplines, particularly chemical biology, to tackle key issues in research field of CM. Therefore, data of current proposal could facilitate a more integrative revealing of mechanism by which components from gardenia possess its activity. Furthermore, strategies of current project would enhance integration of phytochemistry, pharmacology, chemical biology and proteomics in research work of CM, and eventually facilitate confirmation of active components from CM and mechanism by which their biological activities are exerted.

中药治疗或调节人体疾病机制仍然不明，特别是药效物质和作用靶点不清楚，故难以深入揭示其治疗疾病分子机制，严重制约中药现代化国际化。项目基于光偶联探针在中药药效机制研究潜力，及课题组发现中药栀子成分肝保护及肠道菌群调节功效，拟针对栀子体内成分结构特点设计光敏探针，与栀子主要体内成分巧妙对接，并与肝细胞及肠道细菌孵育并经紫外处理，采用LC-MS细致分析样本蛋白组，确定栀子中与肝细胞及肠道菌内蛋白结合化合物，阐明药效靶点。项目基于课题组栀子研究前期数据及学科前沿，高度整合各学科优势，特别是化学生物学领域小分子探针，综合解决中药研究最核心问题，因此，课题研究结果将可能为评价栀子药效机制提供最核心科研数据。项目研究所采用思路与策略，也将有助于中药基础研究广泛采用的天然药化、微生物学、化学生物学和蛋白组学研究的交叉融合，为中药药效基础及药理机制研究提供依据，为解决制约中药现代化国际化提供创新思路。

开展本项目4年多以来，课题组克服疫情影响，上下同心认真组织研究工作，深入开展栀子西红花苷肠道菌功能及神经，抗炎等功效研究课题，揭示西红花苷具显著肠道菌改善功效，数据显示栀子及西红花苷具有肠道菌调节活性潜力，可上调哺乳动物肠道有益菌毛螺菌科，同时下调有害菌梭菌科。同时也首次开创性发现肠道菌也可反作用于西红花苷而使后者在动物肠道内转变为西红花酸，并且阐明该化合物及其葡萄糖醛酸缀合物均为体内药效物质基础，并揭示该代谢物为西红花苷在肠道代谢为西红花酸后，进而被肝脏UGTs酶代谢为西红花酸单葡萄糖醛酸（确定最重要4个UGTs代谢酶），在本领域首次系统阐明该代谢过程的最关键代谢机制。功效方面，体外及整体动物模型研究数据均显示西红花苷具有显著抗炎及改善AD动物药效活性（包括西红花苷及其代谢物西红花酸单葡萄糖醛酸均显示良好活性），同时，我们也结合课题组优势开展产品为导向的功能饮料开发研究，系统研究栀子西红花苷饮料配方，优化方案采用栀子西红花苷配伍菊花和蔗糖，制备出质量高，稳定性好的西红花苷功能饮料。项目这一系列成果数据显示栀子西红花苷具有显著功效价值，拥有相关药物及功能饮料开发良好前景。总体上课题的开展获得重要的栀子西红花苷研究数据及开发思路，揭示栀子西红花苷具有显著抗炎和神经系统功效价值，具有显著食药两用前景优势及相关功能饮料开发良好前景，为当前及今后本领域更全面深入研究开发工作提供重要数据。在项目实施中，共完成并发表学术论文4篇，其中1篇中文核心，3篇SCI收录(其中2篇为中科院1区学术期刊)，并培养研究生研究生研究生研究生7名。