基于功效多向性与谱-效灰色关联分析的中药黄芩药效物质基础辨识与质量控制研究

Efficacy substance identification and quality control is the key and difficult point in Chinese Herbal Medicine (CHM) research. For a long time, the CHM was simply regarded as the same with single compound drugs, and the method used in chemical drug discovering was indiscriminately imitated for digging CHM efficacy substance, in order to find out some single compounds to represent the effect of CHM. However, this method ignored the potential interaction of other ingredient continued in CHM as a whole, did not fit the “multi-component and multi-target” function characteristics of CHM, especially for the of multi-directional efficacy CHM. Whether the chemical ingredients participate in different behavior of CHM is the same? Whether the target for different behavior is the same? Should the same index be used for the quality control of different effect of CHM? These questions have been suspended for a long time in CHM research, and should be clarified forwardly..In this project, Radix Scutellaria (RS) was adopted as model; different samples were collected and prepared, in order to show more chemical representation so could evaluate the sensitive different of chemical and effect under the similar chemical background. LC/MS fingerprints was established, luciferase assay and other biological active assay method was adopted to evaluate the bio effect. Gray correlation was adopted to calculate the grey relational degree. The Fundamental Functional Ingredients (FFI), Specific Functional Ingredients (SFI), Generic Chemical Ingredients (GCI), was distinguished by cluster analysis. Traditional Chinese Medicine syndrome animal model was adopted to verify the clearing heat basic function of RS, and the derivative function such as purging intense heat, eliminating dampness, hemostatic and miscarriage prevention of RS. Serum pharmacochemistry was adopted to verify the different of efficacy substance and targeting generated by different functional tropism, and optimized quality control criterion was established for specific functional behavior respectively. Finally, a novel strategy for efficacy substance identification and quality control of CHM was established in this project, and could be generalize used for enhancing the accuracy and scientificity of CHM.

药效物质辨识和质量控制是中药现代研究的重点和难点，但长期采用单成分分离辨识模式，忽略中药的整体特征，不符合“多成分、多靶点”特点；特别是对具有功效多向性的中药，参与各功效取向的化学物质基础是否相同，作用靶点有无差异，是否应当建立各自不同的质控标准等问题都亟待澄清。本项目以黄芩为研究对象，采集与制备黄芩不同样品，以期在化学背景基本相同的前提下，采用LC/MS指纹图谱、荧光素酶报告基因检测等生物活性检测方法，构建谱-效灰色关联系统，通过“背景求同、因素可控、凸显差异、谱效关联、灰色分析”策略，辨识中药“基础功效成分、导向功效成分、一般化学成分”。采用病证动物模型与血清药物化学方法，验证功效黄芩清热基本功效与“泻火、燥湿、止血、安胎”衍生功效取向差异的科学内涵，证实各功效取向的药效物质基础与作用靶点差异，并建立优化的质控标准；从而构建基于功效多向性与谱-效灰色关联的中药药效物质辨识与质控新模式。

药效物质辨识和质量控制是中药现代研究的重点和难点，但长期采用单成分分离辨识模式，忽略中药的整体特征；特别是对具有功效多向性的中药，参与各功效取向的化学物质基础是否相同，作用靶点有无差异，是否应当建立不同的质控标准等问题都亟待澄清。项目以黄芩为研究对象，构建谱-效灰色关联系统，辨识中药“基础功效成分、导向功效成分、一般化学成分”。验证功效黄芩清热基本功效与“泻火、燥湿、止血、安胎”衍生功效取向差异的科学内涵，并建立优化的质控标准；从而构建基于功效多向性与谱-效灰色关联的中药药效物质辨识与质控新模式。.项目按开展了以下研究：（1）基于网络药理学分析的黄芩抑菌作用（大肠杆菌、金黄色葡萄球菌）药效物质差异分析；（2）基于病证动物模型（高热模型、湿热模型）的黄芩（枯芩、条芩）功效差异取向研究；（3）基于病证动物模型（湿热模型）的黄芩主要有效成分（黄芩苷、黄芩素及其配比）功效取向差异比较研究；（4）基于病证动物模型（肺炎模型、结肠炎模型）的黄芩（枯芩、条芩）功效差异取向及对肠道菌群调节作用差异研究；（5）基于生物活性评价（抑菌、抗炎、促凝）的黄芩（枯芩、条芩及炮制品）功效取向性差异及药效成分地位辨识研究；（6）基于抑菌效应成分指纹谱的黄芩质量控制及标准研究。.项目揭示了中药不同功效取向发挥的“多靶点”作用机制差异；阐明了黄芩不同功效下的基础功效成分、导向功效成分、一般化学成分的差异；建立了基于黄芩抑菌效应的效应成分指纹谱，并建立相关质量标准；上述工作是对中药功效多向性、药效物质基础差异性、质量控制精准性研究的有益探索，为阐明中药多功效物质基础及作用机制差异、实现中药精准用药奠定科学基础，具有较好的科学意义与推广应用价值。.项目执行期间申请专利6项，已授权3项；发表研究论文10篇，其中SCI收录8篇；培养硕士研究12名，毕业4名。项目成果获得湖北省科技进步奖二等奖一项，获得国产保健食品批文1项。