基于目标成分敲除/敲入的中药栀子抗炎药效物质辨识和质量评价方法研究

Quality control of Traditional Chinese Medicines (TCM) is very crucial in modernization and development of TCM, and identification of biologically active components of TCM plays a main role in this process. Although various pharmacological activities of components from gardenia have been revealed by researchers, main active chemicals of gardenia have remained to be investigated. On the basis of target components knock-out/knock-in research mode, efficient separation method and liver-injury pharmacological mode of our previous research, current proposal aims to firstly knock out main components from gardenia, and then comparatively evaluate anti-inflammatory activities of target components, non-target components fraction and crude extract of gardenia in vitro and in vivo. This systematical investigation would reveal which components (iridoid glycosides, crocetin derivatives and other components) of gardenia actually play the key roles to protect inflammation of livers, as well as synergism among these components. Current study would provide crucial data to comprehensively evaluate significance of iridoid glycosides, crocetin derivatives and other components in liver protective effects of gardenia, and therefore, facilitate quality evaluation of gardenia. Furthermore, strategies of current project would enhance integration of separation technology of phytochemistry and pharmacology, and eventually facilitate identification of main active components and quality evaluation of TCM.

中药质量控制是攸关中药现代化发展的重大课题，而中药药效物质辨识则是支撑中药质量控制的重要基石。目前，尽管各国学者在分子生物学水平上揭示了栀子主要成分广泛而显著的药理活性，但栀子关键药效组分仍未阐明。课题采用“目标成分敲除／敲入中药药效成分研究模式”，以前期开发的栀子主要成分高效分离工艺及肝损伤药理模型为基础，采用逆流色谱将栀子中主要成分分别敲除，在体内外水平上分别对比评价目标成分、敲除部位和栀子提取物抗炎药理活性，系统探索栀子关键药效物质究竟是环烯醚萜苷、西红花苷，还是除这两类以外的其它成分(及各成分药理协同作用)。项目研究结果，将为全面评价栀子环烯醚萜苷、西红花苷及其它成分，在栀子抗炎药效作用方面提供科研数据，为栀子质量评价提供依据。项目研究所采用思路、策略与方法，也将有助于中药研究中广泛采用的天然药物化学分离技术手段与药理学研究的交叉融合，为中药药效物质辨识和质量评价思路创新提供依据。

承担本项目4年来，课题组师生认真开展西红花苷抗炎(包括免疫药理)、抗老年痴、体内代谢，以及基于西红花苷最新文献展开的文献综述工作。首先，我们采用小鼠肝炎药理模型和大鼠关节炎模型系统研究和评价栀子抗炎药效成分及其中作用机制，结合栀子环烯醚萜苷和西红花苷含量测定结果，两类成分均可能是栀子抗炎药效物质基础，但环烯醚萜苷是其中主要活性成分，而西红花苷类成分贡献相对小于前者，此外，免疫药理数据也显示西红花苷-1具有显著免疫调节活性。同时，课题组为探索西红花苷体内代谢机制，阐明体内活性成分结构及代谢机制，合成出西红花酸单葡萄糖醛酸，证明该成分存在于血液中，体内体外活性数据显示具有显著抗AD药理活性。此外，课题组还系统评价和总结栀子和西红花中西红花苷分离纯化、分析及构效关系研究文献，并针对性评价西红花苷治疗心血管疾病研究进展，并首次系统总结西红花苷体内药物代谢动力学和制剂学研究现状等工作。总体上，以上工作系统揭示栀子主要药效物质基础，以及西红花苷体内活性及代谢机制，为栀子药材质量评价提供了依据，并为今后西红花苷药物研发抗炎和神经系统药物提供重要前期数据。4年研究工作总体上达到项目预期目标，揭示栀子成分体内药效药理机制，发表SCI论文4篇，中文核心期刊论文4篇，以及大学生创新创业项目奖励2项。培养研究生共9名，已毕业5名。