基于系统药理学方法的苓桂术甘汤治疗NAFLD的关键成分挖掘及协同机制分析

目的：从系统药理学视角结合计算方法，深入解析中药复方苓桂术甘汤（LZD）中的主要有效活性成分对非酒精性脂肪性肝病（nonalcoholic fatty liver disease，NAFLD）的分子作用机制及其协同性。方法：应用TCMSP数据库以及PubChem中检索出LZD的主要化学成分，使用ACD/Labs软件对LZD主要化学成分进行ADMET筛选；从GeneCards、OMIM、DisGeNET等数据库收集NAFLD相关的作用靶点，运用SEA、HitPick靶点预测系统依次检索各活性成分的潜在作用靶点，通过取两者交集获得LZD治疗NAFLD的潜在作用靶点集，同时，提出利用最小集合覆盖算法（SCP）优化出LZD治疗NAFLD的关键成分集；进一步利用STRING数据库获得LZD潜在作用靶点的PPI网络，通过基于网络距离的计算方法发现LZD关键成分之间的协同治疗作用；最后，利用DAVID平台进行GO功能注释和KEGG信号通路富集分析。结果：通过ADMET筛选、靶点预测与识别，筛选出LZD中主要活性成分116个及其作用的78个潜在NAFLD靶点；利用SCP算法优化出LZD中的22个关键成分；对基于STRING构建的PPI网络分析，发现2个核心子模块，同时利用网络扰动计算方法，发现LZD中22个关键成分中有14个成分之间存在协同治疗作用；KEGG通路富集分析，显示LZD主要通过25条相关信号通路，主要涉及：乙型肝炎（hepatitis B）、TNF、丙型肝炎（hepatitis C）、HIF-1等；最后，分子对接验证显示，LZD中的Quercetin、2-methoxyphenylacetone、cinnamic acid、methyl cinnamate、dimethyl phthalate、diethyl phthalate等有效成分与AKT1、CXCL8、JUN、MAPK8、TNF等5个重要靶点均具有较好的结合力。结论：本研究初步揭示了LZD的药效物质基础，探讨了LZD的药理作用机制，为LZD的临床应用以及实验研究提供了重要参考依据。

Excavation of key components and analysis of synergy mechanism for Lingguizhugan Decoction in the treatment of nonalcoholic fatty liver disease based upon systematic pharmacology

OBJECTIVE To employ a perspective of system pharmacology plus calculation methods, molecular mechanism and synergy of major active ingredients in traditional Chinese medicine compound Linggui Zhugan Decoction (LZD) for nonalcoholic fatty liver disease (NAFLD).METHODS The major chemical components of LZD were retrieved from the databases of TCMSP and PubChem. The major chemical components of LZD were screened by ADMET using ACD/Labs software. NAFLD-related target points were collected from the databases of GeneCards, OMIM and DisGeNET, etc. The potential targets of each active ingredient were searched in turn using SEA and HitPick target prediction systems. The potential target set of LZD treatment of NAFLD was obtained by taking an intersection of the two, and at the same time, minimum set covering algorithm (SCP) was proposed for optimizing the key component set of LZD treatment of NAFLD. Protein-protein interaction (PPI) network of potential targets of LZD was obtained through the database of STRING. The synergistic therapeutic effect between key components of LZD was discovered through the calculation method based upon network distance. Finally GO function annotation and KEGG signal pathway enrichment scores were used on the DAVID platform.RESULTS A total of 116 major active ingredients and 78 potential NAFLD targets were detected in LZD through ADMET screening, target prediction and identification. Twenty-two key components in LZD were optimized by SCP algorithm. Two core sub-modules were found based on the PPI network analysis constructed by STRING. And 14/22 key components in LZD had a synergistic therapeutic effect among them by using the network disturbance calculation method. Enrichment analysis of KEGG pathway showed that LZD mainly passed through 25 related signal pathways, mainly involving hepatitis B, tumor necrosis factor (TNF), hepatitis C and HIF-1, etc. Finally molecular docking verification showed that the effective ingredients in LZD (quercetin, 2-methoxyphenylacetone, cinnamic acid, methyl cinnamate, dimethyl phthalate & diethyl phthalate, etc.) had an excellent binding affinity with five important targets (AKT1, CXCL8, JUN, MAPK8 & TNF, etc.).CONCLUSION This study preliminarily revealed the pharmacological material basis of LZD, discussed the pharmacological mechanism of LZD and provided rationales for clinical application and experimental research of LZD.