气滞胃痛颗粒中甘草抗炎镇痛作用的构效组学

Structure-activity Omics on Anti-inflammatory and Analgesic Effect of Glycyrrhizae Radix et Rhizoma in Qizhiweitong Granules

摘要:

目的 采用构效组学的研究方法，阐释甘草发挥抗炎镇痛作用的药效物质。方法 在课题组前期体外筛选的基础上，开展甘草黄酮体内药效研究，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）、Swiss TargetPrediction、疾病相关基因与突变位点数据库（DisGeNET）及在线人类孟德尔遗传数据库（OMIM）等数据库获取甘草黄酮化合物结构及其发挥抗炎镇痛的直接作用靶点；运用检索互作基因（STRING）数据库及Cytoscape 3.7.2软件构建关键蛋白质-蛋白质相互作用（PPI）网络关系；采用分子对接筛选技术对甘草黄酮中度值（degree）高的5个靶点与化学成分分别进行虚拟对接，筛选出关键核心作用靶点，以靶点为桥梁，将甘草中一类或多类化学成分结构与药效紧密关联，根据不同结构类型的甘草黄酮与靶点的亲和力及作用力进一步探讨甘草黄酮化学结构与核心作用靶点的关系。结果 甘草黄酮可降低甲醛致痛大鼠体内前列腺素E₂（PGE₂）含量，有良好的抗炎镇痛效果。获得甘草黄酮发挥抗炎镇痛作用的活性化合物60种，以综合评分为标准，筛选前列腺素内过氧化物合酶2（PTGS2）和促分裂原活化蛋白激酶3（MAPK3）为甘草发挥抗炎镇痛作用的关键核心作用靶点，除黄酮类结构表现出更好的MAPK3选择性外，其余各类甘草黄酮结构均与PTGS2和MAPK3呈现较好结合且含有糖苷片段的结构表现出更好的结合活性。甘草查耳酮类结构中环上引入链状烯烃更利于与靶点结合，甘草黄酮醇中异戊烯基片段可能引起结合活性的差异，黄烷类结构中A环骈合入吡喃环不利于与靶点的结合，异黄酮类化合物B环上取代基的电性、脂溶性和位阻等对结合活性有直接影响。结论 该研究基于甘草黄酮抗炎镇痛药效作用基础上，采用构效组学的研究方法，可以分析甘草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 reveal the pharmacodynamic substances for the anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma by structure-activity omics.Method On the basis of the previous study about the screening of active components in vitro， this study explored the effects of flavonoids in Glycyrrhizae Radix et Rhizoma in vivo. The flavonoids in Glycyrrhizae Radix et Rhizoma and their direct targets for the anti-inflammatory and analgesic effects were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， PharmMapper， Swiss TargetPrediction， DisGeNET， and Online Mendelian Inheritance in Man （OMIM）. STRING and Cytoscape 3.7.2 were employed to establish the protein-protein interaction （PPI） network of key targets. Molecular docking was performed to simulate the binding of five targets with high degrees to flavonoids in Glycyrrhizae Radix et Rhizoma， on the basis of which the key core targets were selected. The targets were used as a bridge to correlate the structures and effects of one or more classes of chemical components in Glycyrrhizae Radix et Rhizoma. According to the binding affinity between flavonoids with different structures in Glycyrrhizae Radix et Rhizoma and targets， the relationships between compound structures and core targets were discussed.Result The flavonoids in Glycyrrhizae Radix et Rhizoma reduced the content of prostaglandin E₂ （PGE₂） in the rat model of pain induced by formalin， demonstrating definite anti-inflammatory and analgesic effects. Sixty active compounds （flavonoids） with anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma were obtained. With the total score as the standard， prostaglandin-endoperoxide synthase 2 （PTGS2） and mitogen-activated protein kinase 3 （MAPK3） were selected as the key core targets of Glycyrrhizae Radix et Rhizoma for the anti-inflammatory and analgesic effects. Except that flavones showed selectivity of binding to MAPK3， the other flavonoids of Glycyrrhizae Radix et Rhizoma showed strong binding to PTGS2 and MAPK3， and the structures containing glycoside fragments showed stronger binding affinity to the targets. The introduction of chain olefins in the ring of chalcones facilitated the binding to the targets. The isopentenyl fragment in flavonols may cause the difference in binding affinity. The parallel combination of a ring into pyran ring in flavanes was not conducive to the binding to the target. The electric charge， liposolubility， and steric hindrance of the substituent group on the B ring of isoflavones directly affected the binding affinity.Conclusion This study adopts structure-activity omics to analyze the material basis for the anti-inflammatory and analgesic effects of Glycyrrhizae Radix et Rhizoma. Structure-activity omics provides new ideas and methods for predicting the pharmacodynamic substances of traditional Chinese medicine.