基于化学信息学的抗肝纤维化中药药效物质组与靶标群相互作用分析方法学研究

The characterization of interaction between active components of TCM formula and multiple targets is the key for evaluating their safety and effects and elucidating their mechanism. Now, there is a lack of idea for revealing the interaction between “multi-component and multi-target” in TCM formula with anti-liver fibrosis effect quickly and efficiently. In this project, we aim to develop a systematic cheminformatics analytical method based on interaction of multi-component and multi-target, composed of three aspects: virtual screening, construction of liver fibrosis related Chinese herbal visual network pharmacology platform and experimental validation of interaction between components and targets. S100A4 were taken as examples to use above systematic analytical method. A liver fibrosis related TCM formulae, herbs and components database is firstly constructed as a research basis, then we conduct a virtual screening based method to identify herbal components targeting S100A4 with anti-liver fibrosis effect from this database, experiments are performed to verify predicted results. Meanwhile, we construct a liver fibrosis related Chinese herbal visual network pharmacology platform, and select Yin chen hao tang and Fu zheng hua yu fang to showcase the function of platform in investigating action mechanisms of TCM formula. In vitro and in vivo experiments are conducted to validate the “components-targets-pathways” network, and finally demonstrating the practicability and reliability of platform. This project will provide new thoughts and technologies for the screening of active components in TCM and elucidating their complex molecular mechanism.

明确中药复方体内药效物质组与靶标群的相关效应，是评价安全有效性和探讨药效机制的关键。针对尚缺少快速高效地表征抗肝纤维化中药复方“药效活性成分-靶标群”方法的现状，本项目拟构建以抗肝纤维化中药复方“药效物质组-靶标群”相互作用为核心，由药效物质组的虚拟筛选、肝纤维化相关中草药可视化网络药理平台构建及应用、药效物质组-靶标群验证三个方面组成的中药化学信息学分析方法体系。首先以钙结合蛋白S100A4为示范，构建肝纤维化相关中药复方化学成分库，虚拟筛选靶向的抗肝纤维化中药活性单体，并进行体外验证，建立中药药效活性成分虚拟筛选体系。同时，构建肝纤维化相关中草药可视化网络药理平台，以茵陈蒿汤和扶正化瘀方为例展示平台在抗肝纤维化中药复方机制探讨方面的应用，并通过物理学和药理学验证得到的成分-靶标-通路网络，证明平台实用性及可靠性，从而为中药复方药效物质基础及其分子机制的科学阐释提供新思路和新技术。

明确中药复方体内药效物质组与靶标群的相关效应，是评价安全有效性和探讨药效机制的关键。本项目经过四年的研究，基本按计划完成项目任务书中制定的主要研究内容。我们首先利用UPLC/Q-TOF/MS技术对茵陈蒿汤和扶正化瘀方的化学物质组进行了研究。然后，基于比较药动学和文本挖掘对茵陈蒿汤和扶正化瘀方抗肝纤维化潜在药效物质基础进行了探讨，结果显示，茵陈蒿汤中有18个潜在活性成分，扶正化瘀方则有40个潜在活性成分。此外，我们还构建了肝纤维化相关中草药网络药理平台，对茵陈蒿汤和扶正化瘀方的靶标群进行了鉴别，探讨了其潜在药理作用机制。最后，药理验证结果表明，扶正化瘀方通过抑制肝星状细胞活化发挥抗肝纤维化机制的主要活性成分为丹参脂溶性部位、五味子酯甲、五味子乙素、丹酚酸A、丹酚酸B、丹参酮II-A、虫草素、二氢丹参酮I和山奈酚。其中，丹参脂溶性部位通过调控活化肝星状细胞中的JAK1/STAT3信号通路，抑制肝星状细胞的活化，进而抗肝纤维化；山奈酚、丹酚酸A和五味子乙素可与PPARG直接结合，发挥抗肝纤维化作用；二氢丹参酮I则通过靶向P53和STAT3，进而抑制肝星状细胞的活化，发挥体内外抗肝纤维化活性。本项目的实施为中药复方药效物质基础及其分子机制的科学阐释提供了新思路。在本基金的资助下，已培养硕士研究生3名，发表SCI 论文4篇，核心期刊3篇，其中第一标注5篇，第二标注2篇；申请国家发明专利1项。