基于网络药理学探讨苍附导痰...征大鼠胰岛素抵抗的作用机制_华宙佳.pdf

中国现代应用药学 2023 年 2 月第 40 卷第 4 期 Chin J Mod Appl Pharm,2023 February,Vol.40 No.4 461 基于网络药理学探讨苍附导痰汤加减方对多囊卵巢综合征大鼠胰岛素抵抗的作用机制 华宙佳，王晨晔，俞佳，丁彩飞*(浙江省中西医结合医院生殖科，杭州 310003)摘要：目的 基于网络药理学探讨苍附导痰汤加减方(modified Cangfu Daotan decoction，CFDTD)对多囊卵巢综合征(polycystic ovarian syndrome，PCOS)大鼠胰岛素抵抗(insulin resistance，IR)的影响。方法 利用 TCMSP、TCM DatabaseTaiwan、Drugbank、GeneCards 数据库检索 CFDTD 中的活性化合物并筛选作用靶点；从 TTD、PubMed 及PharmGKB 数据库检索与 PCOS 相关的疾病靶点，成分靶点与疾病靶点映射后使用 Cytoscape 3.7.1 软件构建药物有效成分-靶点蛋白相互作用网络。然后，采用 STRING 和 Cytoscape 软件构建蛋白相互作用网络并筛选核心靶点。随后，利用Cytoscape 软件中的 ClueGO 插件进行 Kyoto Encyclopedia of Genes and Genomes(KEGG)通路分析。最后，采用来曲唑联合高糖高脂饮食构建 PCOS 伴 IR 大鼠模型加以验证；给药组分别给予 15，30 gkg1d1 CFDTD 及 50 mgkg1d1盐酸二甲双胍(metformin hydrochloride，MH)灌胃；对照组和 PCOS 组给予等体积生理盐水灌胃，干预 4 周。全自动化学分析仪检测大鼠空腹血糖(fasting blood glucose，FPG)、空腹胰岛素(fasting insulin，FINS)、促卵泡生长激素(follicle stimulating hormone，FSH)、促黄体生成素(luteinizing hormone，LH)、睾酮(testosterone，T)、雌二醇(estradiol，E2)含量差异并计算胰岛素抵抗指数(homeostatic model assessment insulin resistance index，HOMA-IR)。HE 染色观察大鼠卵巢组织病理变化。Western blotting 检测大鼠卵巢组织胰岛素受体(insulin receptor，INSR)、胰岛素样生长因子-I(insulin-like growth factor-I，IGF-I)、胰岛素样生长因子 I 受体(insulin-like growth factor-I receptor，IGF-IR)、胰岛素(insulin，INS)、肿瘤坏死因子-(tumor necrosis factor-，TNF-)、白介素-6(interleukin-6，IL-6)蛋白表达水平。结果 CFDTD 筛选得到 86 个活性成分及 126 个CFDTD 治疗 PCOS 潜在相关靶点。KEGG分析结果显示，潜在靶点主要涉及 insulin resistance，steroid hormone biosynthesis，ovarian steroidogenesis，estrogen signaling pathway，PI3K-Akt signaling pathway，type II diabetes mellitus 等信号通路。蛋白相互作用分析提示 INS、TNF-及 IL-6 为蛋白相互作用网络中的关键靶点。与 PCOS 组相比，给药组大鼠血清 FPG、FINS、HOMA-IR、LH、T、E2的含量明显减少(P 均0.01)及卵巢组织形态明显恢复。Western blotting 检测结果显示 CFDTD显著降低 PCOS 大鼠卵巢组织 INSR、IGF-IR、INS、TNF-及 IL-6 蛋白表达。结论 CFDTD 可能通过下调 INSR、IGF-IR、INS、TNF-及 IL-6 蛋白表达，改善 PCOS 大鼠 IR。关键词：多囊卵巢综合征；苍附导痰汤加减方；网络药理学；胰岛素抵抗；生殖激素 中图分类号：R285.5 文献标志码：A 文章编号：1007-7693(2023)04-0461-10 DOI:10.13748/ki.issn1007-7693.2023.04.005 引用本文：华宙佳,王晨晔,俞佳,等.基于网络药理学探讨苍附导痰汤加减方对多囊卵巢综合征大鼠胰岛素抵抗的作用机制J.中国现代应用药学,2023,40(4):461-470.Investigation on Mechanism of Action of Modified Cangfu Daotan Decoction on Insulin Resistance in Polycystic Ovarian Syndrome Rats Based on Network Pharmacology HUA Zhoujia,WANG Chenye,YU Jia,DING Caifei*(Department of Reproductive,Zhejiang Chinese Medicine and Western Medicine Integrated Hospital,Hangzhou 310003,China)ABSTRACT:OBJECTIVE To explore the effects of modified Cangfu Daotan decoction(CFDTD)on insulin resistance(IR)in polycystic ovarian syndrome(PCOS)rats based on network pharmacology analysis.METHODS TCMSP,TCM databaseTaiwan,Drugbank and GeneCards databases were used to screen out the active chemical components and action targets of CFDTD.TTD,PubMed,and PharmGKB databases were used to screen out the disease targets associated with PCOS.After mapping the component target with the disease targets,the Cytoscape 3.7.1 software was used to construct the component-target interaction network of CFDTD.Next,STRING and Cytoscape software was used to construct protein-protein interaction network and identify the hub targets.Subsequently,Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis was performed by using the ClueGO plug-in tool in Cytoscape.Furthermore,experimental validation was conducted in the PCOS with IR model rats induced by letrozole combined with high-fat and high-sugar diet.The treated groups were given CFDTD at a 基金项目：杭州市卫生科技计划项目(OO20190089)作者简介：华宙佳，女，硕士，主治医师 E-mail: *通信作者：丁彩飞，女，主任医师 E-mail: 462 Chin J Mod Appl Pharm,2023 February,Vol.40 No.4 中国现代应用药学 2023 年 2 月第 40 卷第 4 期 dose of 15 and 30 gkg1d1,and metformin hydrochloride(MH,50 mgkg1d1)respectively,the control and PCOS groups were given equivoluminal normal saline for 4 weeks.The serum levels of fasting blood glucose(FPG),fasting insulin(FINS),follicle stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T)and estradiol(E2)of rats were detected by automatic chemical analyzer,and the homeostatic model assessment insulin resistance index(HOMA-IR)was then calculated.HE staining was used to observe the morphological changes of ovarian tissues.The protein expression levels of insulin receptor(INSR),insulin-like growth factor-I(IGF-I),insulin-like growth factor-I receptor(IGF-IR),insulin(INS),tumor necrosis factor-(TNF-),interleukin-6(IL-6)were detected by Western blotting.RESULTS The 86 active components of CFDTD and 126 potential targets related to PCOS were screened by network pharmacology analysis.KEGG analysis showed that the potential targets were closely related to signaling pathway such as insulin resistance,steroid hormone biosynthesis,ovarian steroidogenesis,estrogen signaling pathway,PI3K-Akt signaling pathway,type II diabetes mellitus,etc.Protein interaction analysis indicated that INS,TNF-,and IL-6 were the hub targets of protein interaction network.Compared with the PCOS group,the levels of FPG,FINS,HOMA-IR,LH,T and E2 in the treated groups decreased significantly(P all0.01),and the damaged degree of ovarian tissue was significantly reduced.Western blotting showed that CFDTD could significa