基于网络多靶标的中药复方分析方法学研究

In terms of the current situation where it still lacks integrated analysis methodology in macroscopic and microcosmic view on TCM formula, this study exemplified Sini decoction (SND), which not only applied several tools and approaches based on multidisciplinary but also combined analyses on integrity and molecular level. Taking the protein expression and metabolite regulation of the essential characteristics of the disease and syndrome as network multi-target, we studied the relationship between the SND compatibility and the regulation pattern of network multi-target and then analyzed the contribution and relationship of the various medicinal herbs and their compatibility to SND. From the perspective of the network multi-target regulation, we dissected the mechanisms of SND on myocardial infarction and then established the target network of SND. The effective components and its corresponding potential targets were virtually screening by the binding capacity between PK-marker of SND and target which was calculated by the method of network pharmacology. Then, the relationship between the effective compounds and targets was validated in cell model. By establishing the related network of "SND-herb-multi-components-multi-targets", we explained the effective components of SND and scientific connotation of SND compatibility. We established analysis methodologies for TCM based on network multi-targets, which provide new methodological supports for the modern molecular study of TCM formula. It will certainly be a great significance for scientific value of TCM formula quality as well as the research and development of new medicine based on network regulation.

针对中药复方分析仍然缺少宏观和微观相结合的分析方法学的现状，本研究以四逆汤为研究示范，综合运用多学科手段和工具，将整体分析与分子水平分析相结合，以反映病证本质特征的蛋白表达和代谢物调控为网络多靶标，研究全方和不同配伍组合与网络多靶标调控模式的相关性，辨析各药材及其配伍对全方整合调节效应的贡献度和关联关系，从网络多靶标调控的角度诠释四逆汤治疗心肌梗死的现代生物学意义，探索构建四逆汤赖以发挥作用的靶点网络，通过分子模拟方法计算预测四逆汤药代标示物与靶点的结合能力，虚拟筛选中药复方的药效成分及其可能靶标，并在细胞模型上验证其对靶标的活性，构建"四逆汤-药材-多成分-多靶点"相关网络，揭示四逆汤药效物质基础和配伍的科学内涵。通过本项目的研究，构建基于网络多靶标的中药复方分析方法学，为中药复方的分子水平分析提供新方法，这对解决中药复方质量评价的科学性和基于网络调控的新药研发具有重要意义。

本研究以四逆汤为研究示范，综合运用多学科手段和工具，将整体分析与分子水平分析相结合，建立了“四逆汤-药材-多成分-多靶点”相关网络，筛选鉴定了四逆汤体内化学物质组成分；以正常和病理的细胞膜色谱模型筛选了附子及四逆汤中诱导心衰的活性组分；阐明四逆汤主要通过影响琥珀酸合成反应，参与NADH的生物合成过程，促进心肌收缩，维持细胞骨架和活力，参与胶原合成反应、转醛酶反应、能量转运与代谢反应、柠檬酸合成反应、3-羟基异丁基辅酶A合成酶、乙酰辅酶A脱氢酶等反应，调节离子通道活性来发挥保护心肌细胞、治疗心肌梗死的作用；揭示了附子总生物碱是主要的活性成分，而干姜总姜酚和甘草总黄酮总皂苷是辅助成分，从而阐明中药复方“君臣佐使”的配伍规律和协同拮抗的配伍机制。 本项目的研究不仅为揭示四逆汤药效物质基础和配伍的科学内涵提供理论依据， 也为其它中药复方的分子水平分析提供新方法，这对解决中药复方质量评价的科学性和基于网络调控的新药研发具有重要意义。项目研究发表SCI 论文１９ 篇，培养研究生７ 名，超额完成任务指标。