利用单克隆抗体特异性敲除技术解析栀子苷与栀子药效关联性的研究

The correlation between the representative ingredients and the efficacy of herb medicine is one of the key scientific issues to explain the material basis of Chinese medicines.Because the pharmacodynamic efficacy of Gardenia was relatively clear,it was selected for the object in this study. We use geniposide monoclonal antibody as a tool to establish the specific immune affinity knockout technology methods.The efficacies of cardiovascular protection, cholagogic and liver protection, nerve protection and antipyretic analgesic of Gardenia water extract were compared before knockout, after knockout and also after add a certain amount of geniposide, by this way we can clarify the influence of the content of the geniposide on the effects of gardenia.With high sensitivity and specificity immunoassay methods which are suitable for the detection of microscale amounts of biological samples,the contribution of geniposide to the efficacy of gardenia and the relatationship between content-efficacy-location-pathway were analyzed with the classic model. The standard of evaluation system was a combination of cell and entirety,the static state and the dynamic state, and the efficacy and factors. This study will provide a powerful experiment evidence for the quality control, metabolism, distribution, and pharmacodynamic analysis of gardenia herbs. This will also offering a new approach,technology and methods for the research of Pharmacology efficacy studies of chinese herb medicine.

代表性成分与药物功效之间的相关性，是解析中药药效物质基础的关键科学问题之一。本研究选择药效相对明确的栀子为对象，利用栀子苷单克隆抗体为工具，建立特异性免疫亲和敲除技术方法，通过对比敲除栀子苷前后栀子水提物药理药效的异同，阐明栀子苷与栀子功用主治的相关性，并在此基础上采用定量回添栀子苷的方法，探讨栀子苷不同含量对栀子抗炎镇痛、保肝利胆、神经保护的方面药效的影响。研究借助灵敏度高、特异性强、适宜微量生物样品检测的免疫分析方法，利用与栀子传统主治功效相对应的经典药理学评价模型，以整体与细胞、静态与动态、药效与生物学指标相结合的评价体系为标准，阐明栀子苷对栀子药效的贡献度，解析二者在效-量-径-位方面的相互关系。该研究将为栀子药材的质量控制、代谢分布及药效分析提供有力的实验依据，并为中药药理药效研究开辟新的思路，提供新的技术与方法。

代表性成分与药物功效之间的相关性，是解析中药药效物质基础的关键科学问题之一。本研究选择药效相对明确的栀子为对象，建立特异性免疫亲和敲除技术方法，敲除栀子水提液中单个小分子成分栀子苷，敲除率最高可达71.16%，平均敲除率约为50%。栀子苷与栀子均具有抗炎、保肝、血管内皮保护、神经保护作用，通过对比敲除栀子苷前后栀子水提液药效的异同，并在此基础上采用定量回添栀子苷的方法研究栀子苷对栀子药效的贡献度，发现栀子苷是栀子水提液发挥神经保护作用的主要成分，而在抗炎、保肝、血管内皮保护方面，栀子苷并不是栀子水提液发挥药效的唯一有效组分。本研究为栀子药材的质量控制提供依据，同时为中药药效物质基础研究建立了新模式。