基于网络药理学及分子对接技术探究清肝解郁汤加减治疗多囊卵巢综合征的作用机制

Exploring the Mechanism of Action of Qinggan Jieyu Decoction and Its Modified Formula in the Treatment of Polycystic Ovary Syndrome Based on Network Pharmacology and Molecular Docking Technology

摘要: 目的：通过网络药理学探究中药复方清肝解郁汤加减治疗多囊卵巢综合征的有效分子机制，并通过分子对接技术加以验证。方法：运用TCSMP数据库筛选出清肝解郁汤加减中各中药的活性成分，再用Pubchem及SwissTarget数据库筛选出每个活性成分所对应的靶点，运用Genecards、TTD、Drungbank、OMIM等数据库收集多囊卵巢综合征的疾病靶点；将清肝解郁汤加减的活性成分对应的靶点与多囊卵巢综合征的疾病靶点取交集，作为关键靶点；运用STRING数据库构建蛋白与蛋白互作网络(PPI)；再用Cytoscape3.10.1软件构建药物–活性成分–潜在靶点网络；运用DAVID数据库进行GO以及KEGG富集通路的分析；最后应用Autodock Tools及Autodock Vina对中药复方中药物的核心成分与疾病的关键靶点进行分子对接验证，最后运用Pymol进行可视化分析。结果：获得清肝解郁汤加减的活性成分所对应的靶点1054个，多囊卵巢综合征疾病靶点1189个，取交集得到清肝解郁汤加减与多囊卵巢综合征的关键靶点175个；用PPI网络对关键靶点进行分析，根据Degree值大小得到核心靶点4个，分别为甘油醛-3-磷酸脱氢酶(GAPDH)、丝氨酸/苏氨酸蛋白激酶(AKT1)、白细胞介素6 (IL-6)、肿瘤坏死因子(TNF)；富集分析结果显示，清肝解郁汤加减可能通过调节缺氧诱导因子1信号通路(HIF-1 signaling pathway)、磷脂酰肌醇3-激酶-蛋白激酶B信号通路(P13K-Akt signaling pathway)、叉头框O类家族成员蛋白(FoxO signaling pathway)等信号通路来改善多囊卵巢综合征。分子对接结果显示，核心化合物槲皮素与GAPDH、TNF，β-谷甾醇与TNF，山柰酚与GAPDH，黄芩素与GAPDH等有良好的结合活性。结论：清肝解郁汤加减可通过多靶点、多通路发挥治疗多囊卵巢综合征的作用，为多囊卵巢综合征的治疗及新药研究提供了新思路。

Abstract: Objective: Exploring the effective molecular mechanism of the addition of the Chinese herbal compound Qinggan Jieyu Decoction and its modified formula for the treatment of polycystic ovary syndrome by network pharmacology and verifying it by molecular docking technique. Methods: Screening of the active ingredients of each Chinese medicine in Chinese medicine compound formulae using the TCSMP database. Then, the Pubchem and SwissTarget databases were used to screen the targets corresponding to each active ingredient. Databases such as Genecards, TTD, Drungbank, and OMIM were used to collect the disease targets of PCOS. The intersection of the targets corresponding to the active ingredients of Qinggan Jieyu Decoction and its modified formula and the disease targets of PCOS was taken as the key targets. Construct protein-protein interaction (PPI) network using STRING database; then construct drug-active ingredient-potential target network using Cytoscape 3.10.1 software. The DAVID website was used for GO analysis and KEGG enrichment analysis. Finally, software such as Pymol and Autodock Tools were applied to verify the molecular docking of the core components and key targets, and Pymol was used for visual analysis. Results: A total of 1054 targets corresponding to the active ingredients of QingganJieyu Decoction and its modified formula were obtained, and 1189 disease targets of PCOS were obtained. By taking the intersection, 175 key targets between Qinggan Jieyu Decoction and its modified formula and PCOS were obtained. The key targets were analyzed using the PPI network. According to the Degree values, 4 core targets were obtained, namely glyceraldehyde-3-phosphate dehydrogenase (GAPDH), serine/threonine protein kinase (AKT1), interleukin-6 (IL-6), and tumor necrosis factor (TNF). The results of the enrichment analysis showed that Qinggan Jieyu Decoction and its modified formula may improve PCOS by modulating signalling pathways such as the hypoxia-inducible factor 1 signalling pathway (HIF-1 signalling pathway), phosphatidylinositol 3-kinase-protein kinase B signalling pathway(P13K-Akt signalling pathway), and the FoxO signalling pathway. The results of molecular docking showed that the core compound quercetin had good binding activities with GAPDH and TNF, β-sitosterol with TNF, kaempferol with GAPDH, and baicalein with GAPDH. Conclusion: Qinggan Jieyu Decoction and its modified formula can play a role in treating PCOS through multiple targets and multiple pathways, providing new ideas for the research of new drugs for treating PCOS.