南方医科大学学报

南方医科大学学报 ›› 2022, Vol. 42 ›› Issue (1): 1-12.doi: 10.12122/j.issn.1673-4254.2022.01.01

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补肾化痰方治疗多囊卵巢综合征的作用机制：基于网络药理学和分子对接方法

高梦雅，洪艳丽，崔媚婷，黄菁宇，谈 勇，聂晓伟   

  1. 南京中医药大学附属医院，江苏 南京 210029
  • 出版日期:2022-01-20 发布日期:2022-03-02

Bushen Huatan recipe for treatment of polycystic ovary syndrome: therapeutic mechanism based on network pharmacology and molecular docking

GAO Mengya, HONG Yanli, CUI Meiting, HUANG Jingyu, TAN Yong, NIE Xiaowei   

  1. Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China
  • Online:2022-01-20 Published:2022-03-02

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摘要： 目的 探究补肾化痰方治疗多囊卵巢综合征的主要活性成分、靶点、通路等药理学作用机制。方法 通过中药系统药理学数据库与分析平台（TCMSP）筛选出补肾化痰方中药物的主要活性成分，使用 PubChem和Swiss Target Prediction预测活性成分相关作用靶点；应用GeneCards、OMIM筛选治疗疾病多囊卵巢综合征的作用靶点；将药物靶点与疾病靶点通过Uniprot校正后，取交集靶点；结合STRING构建蛋白-蛋白互作（PPI）网络，并利用Cytoscape3.7.2中的CytoNCA插件对交集靶点进行PPI分析，筛选出核心靶点；采用DAVID数据库进行基因功能GO富集分析和KEGG通路富集分析；利用AutoDock对补肾化痰方中的核心活性成分和核心靶点进行分子对接验证。以动物实验进行验证，24只SPF级雌性C57BL/6J小鼠按随机数字表法分为3 组：对照组［n=8，1 mg/（kg?d）生理盐水灌服35 d］，模型组［n=8，1 mg/（kg?d）来曲唑溶于1%羧甲基纤维素（CMC）中，连续灌服35 d，并予高脂饲料喂养］，治疗组［n=8，造模后予以1 mg/（kg?d）补肾化痰方颗粒剂溶于5%生理盐水中，连续灌服35 d］。小鼠末次给药后，取卵巢组织于-80 ℃保存，Western blot法检测小鼠卵巢组织中核心靶点和核心通路的表达。结果 补肾化痰方中14味中药共筛选出125个潜在活性成分，990个药物靶点，4759个PCOS靶点，药物与PCOS交集靶点434个；核心活性成分主要为β-谷甾醇、山奈酚、槲皮素等，核心靶点主要为PIK3CA、PIK3R1、APP、AKT1、MAPK1等；GO功能富集主要包括药物反应、凋亡过程的负调控、RNA聚合酶Ⅱ启动子转录的正调控等；KEGG富集通路主要包括癌症通路、HIF-1信号通路、Rap1信号通路、PI3K-Akt信号通路等；分子对接结果显示核心活性成分β-谷甾醇、山奈酚、槲皮素与核心靶点PIK3CA、PIK3R1、APP、AKT1、MAPK1均有强烈的结合能力；动物实验结果表明，补肾化痰方能明显改善多囊卵巢综合征相关症状，并下调PI3K，AKT蛋白表达，上调MAPK1蛋白表达，验证了网络药理学的部分预测结果。结论 揭示了补肾化痰方多成分、多靶点、多途径的作用特点，预测并通过实验初步验证了补肾化痰方治疗多囊卵巢综合征的可能作用机制。

关键词: 多囊卵巢综合征；补肾化痰方；网络药理学；分子对接；作用机制

Abstract: Objective To explore the pharmacological mechanism of Bushen Huatan (BSHT) recipe in the treatment of polycystic ovary syndrome (PCOS). Methods The active ingredients in the component drugs of the recipe were screened through TCMSP, and their potential targets were predicted by PubChem and Swiss target prediction. Genecards and OMIM were used to screen the therapeutic targets in the treatment of PCOS. The drug targets and disease targets were corrected using Uniprot, and the intersection targets were obtained. The protein-protein interaction (PPI) network was constructed using STRING, and the intersection targets were analyzed with CytoNCA to screen the core targets. DAVID was used for GO enrichment analysis and KEGG pathway enrichment analysis, and the core components and core targets were verified using AutoDock. Animal experiment was performed to verify the results using a female C57BL/6J mouse model of PCOS, treated daily with 1 mg/kg BSHT recipe granule for 35 days, and the ovarian expressions of the core targets and pathways were detected using Western blotting.Results We identified a total of 125 potential active ingredients from the 14 component drugs in the recipe, 990 drug targets, 4759 PCOS targets and 434 intersection targets. The core active ingredients of the recipe included β -Sitosterol, kaempferol, and quercetin, whose core targets included PIK3CA, PIK3R1, APP, AKT1, and MAPK1. GO enrichment analysis highlighted such processes as drug reaction, negative regulation of apoptosis, and positive regulation of transcription from RNA polymerase II promoter. The enriched KEGG pathways included primarily the cancer pathway and PI3K-Akt signaling pathway. Molecular docking showed that the core active ingredients had strong binding ability with the core targets. In the animal experiment, BSHT recipe was shown to improve the symptoms, down-regulate the expressions of PI3K and Akt proteins and up-regulate MAPK1 expression in the ovary of mice with PCOS. Conclusion The therapeutic mechanism of BSHT recipe for PCOS involves multiple active ingredients, multiple therapeutic targets and multiple pathways.

Key words: polycystic ovary syndrome; Bushen Huatan recipe; network pharmacology; molecular docking; mechanism