气滞胃痛颗粒中香附抗炎镇痛作用的构效组学

Structure-activity Omics on Anti-inflammatory and Analgesic Effect of Cyperi Rhizoma in Qizhi Weitong Granules

摘要:

目的 采用构效组学的研究方法，阐释香附发挥抗炎镇痛作用的药效物质。方法 基于课题组前期体外药效筛选，开展香附黄酮组分体内药效研究，并通过中药系统药理数据库和分析平台（TCMSP）、药效预测靶点数据库（PharmMapper）和Swiss TargetPrediction等数据库及文献调研获取香附黄酮类成分及作用靶点，采用疾病相关基因与突变位点数据库（DisGeNET）和人类孟德尔遗传数据库（OMIM）等数据库收集抗炎镇痛靶点，取交集靶点作为香附黄酮抗炎镇痛的直接作用靶点，构建核心靶点蛋白质-蛋白质相互作用（PPI）网络。基于构效组学的研究方法，以靶点为桥梁，将香附中一类或多类化学成分结构与药效紧密关联，将化学成分按照结构分类，并通过SYBYL-X 2.1.1、PyMOL及Discovery Studio 4.5 visualizer软件将成分与核心靶点进行分子对接，选取整体对接活性最好的2个靶点，探讨化合物结构与靶点的关系。结果 香附黄酮对小鼠甲醛致痛模型具有良好的抗炎镇痛作用，筛选获得香附黄酮成分18个和直接作用靶点115个，分析得到高活性的核心靶点为蛋白激酶B1（Akt1）、白细胞介素-1β（IL-1β）、细胞肿瘤抗原p53（TP53）、前列腺素内过氧化物合酶2（PTGS2）和基质金属蛋白酶-9（MMP-9）。香附黄酮成分按照结构类型分为双黄酮类、黄酮醇类、黄酮类及黄烷类，经分子对接筛选，成分与TP53和PTGS2整体对接活性最好。构效组学研究结果表明，双黄酮类结构是香附黄酮中整体与靶点结合最好的结构，但其多羟基醚化带来与PTGS2结合活性的明显下降；糖苷与PTGS2有更好的结合，而黄酮醇的A环引入长链烃基则与TP53结合更佳，B环取代基的变化不是影响结合活性的主要因素；3，4-二羟基黄烷结构与TP53的结合活性优于3-羟基黄烷，但与PTGS2的结合未表现出优势。结论 该研究基于香附黄酮抗炎镇痛的药效作用基础上，采用构效组学研究方法，可以分析香附黄酮抗炎镇痛的物质基础，同时构效组学为中药药效物质的阐释提供新思路新方法。

Abstract:

Objective To elucidate the pharmacodynamic substances responsible for the anti-inflammatory and analgesic effects of Cyperi Rhizoma by structure-activity omics.Method On the basis of the previous in vitro efficacy study by our research group， this study explored the in vivo efficacy of the flavonoids in Cyperi Rhizoma. The flavonoids in Cyperi Rhizoma and their targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， PharmMapper， Swiss TargetPrediction， and available articles. The targets of the anti-inflammatory and analgesic effects were collected from DisGeNET and Online Mendelian Inheritance in Man （OMIM）. The common targets shared by flavonoids and the effects were selected as the direct targets of flavonoids endowing Cyperi Rhizoma with anti-inflammatory and analgesic effects， and protein-protein interaction （PPI） network of the core targets was constructed. The method of structure-activity omics was employed to correlate the structure and efficacy of one or more classes of chemical components in Cyperi Rhizoma with the targets as a bridge. The components were classified according to structure. Molecular docking of components to core targets was carried out via SYBYL-X 2.1.1， PyMol， and Discovery Studio 4.5 visualizer. Two targets with the highest binding affinity were selected to explore the relationship between compound structures and targets.Result The flavonoids in Cyperi Rhizoma exerted anti-inflammatory and analgesic effects on the mouse model of pain induced by formaldehyde. Eighteen components and 115 direct targets were screened out， and the core targets with high activities were protein kinase B1 （Akt1）， interleukin-1β （IL-1β）， cellular tumor antigen p53 （TP53）， prostaglandin-endoperoxide synthase 2 （PTGS2）， and matrix metalloproteinase-9 （MMP-9）. According to the structures， the flavonoids in Cyperi Rhizoma were classified into bioflavonoids， flavonols， flavones， and flavanes. The molecular docking results showed that flavonoids of Cyperi Rhizoma had the highest binding affinity to TP53 and PTGS2. The results of structure-activity omics showed that bioflavonoids represented the best binding structure to the targets， while their polyhydroxyl etherification resulted in a significant decrease in the binding affinity to PTGS2. Glycosides had higher binding affinity to PTGS2. The introduction of the long-chain hydrocarbon group to the A ring of flavonols facilitated the binding to TP53， while the change of B ring substituents was not the main factor affecting the binding affinity. The 3，4-dihydroxyl flavane outperformed 3-hydroxyl flavane in the binding to TP53， while the two compounds showed similar binding affinity to PTGS2.Conclusion The method of structure-activity omics was used to analyze the material basis for the anti-inflammatory and analgesic effects of flavonoids in Cyperi Rhizoma. Structure-activity omics provides new ideas for revealing the pharmacodynamic substances of traditional Chinese medicine.