网络药理学整合巨噬细胞差异基因揭示双氢杨梅树皮素改善急性肺损伤的作用机制及实验验证  

Network pharmacology integrates the differential genes of macrophages to explain the mechanism of ampelopsin alleviating acute lung injury and experimental verification

作　　者：唐木兰 曾春晖[1] 黄鑫波 王溢 朱海滨 杨柯[1] TANG Mu-lan;ZENG Chun-hui;HUANG Xin-bo;WANG Yi;ZHU Hai-bin;YANG Ke(Guangxi University of Chinese Medicine,Nanning 530000)

机构地区：[1]广西中医药大学,南宁530000

出　　处：《中南药学》2023年第5期1164-1169,共6页Central South Pharmacy

基　　金：广西中医药大学2021年校级硕士研究生科研创新项目(No.YCXJ2021014);广西自然科学基金项目(No.2020GXNSFDA238028);国家中医药管理局“中药炮制传承技术传承基地”资质项目(桂中医药发[2020]9号);广西中医药大学2020年校级硕士研究生科研创新项目(No.YCSZ2020009);广西教育厅广西研究生教育创新计划资助项目(No.JGY2014086)。

摘　　要：目的基于网络药理学和巨噬细胞差异表达基因探讨双氢杨梅树皮素改善急性肺损伤的作用机制及实验验证。方法应用Cytoscape软件,通过PharmMapper服务器、GeneCards和OMIM数据库构建“化合物-靶点-通路”网络;利用STRING数据库构建靶点蛋白互作网络,推测核心靶点;使用Metascape数据库对核心靶点进行GO、KEGG富集分析。下载巨噬细胞芯片数据,利用GEO2R分析芯片数据的差异表达基因,DAVID平台进行差异基因KEGG通路富集。采用流式细胞术检测M1型和M2型巨噬细胞标志物F4/80、CD11b、CD86和CD206的表达;采用Western blot法检测肺组织中PI3K和Akt蛋白表达量。结果筛选出102个双氢杨梅树皮素抗急性肺损伤的潜在靶点,主要富集在PI3K/Akt信号通路、JAK/STAT信号通路、雌激素信号通路等;分子对接结果显示双氢杨梅树皮素与MMP9和IGF1具有较好的结合活性。差异表达基因富集到PI3K-Akt、细胞因子-细胞因子-受体相互作用、趋化因子等炎症相关信号通路。流式细胞术实验证明,双氢杨梅树皮素能明显增加M2型巨噬细胞极化和降低M1/M2的比例,且明显降低肺组织中PI3K和Akt蛋白表达量。结论通过网络药理学和动物实验证实双氢杨梅树皮素通过PI3K/Akt信号通路调节巨噬细胞极化改善急性肺损伤,为进一步阐明双氢杨梅树皮素改善ALI/ARDS的机制提供理论依据。Objective To determine the mechanism of ampelopsin in the treatment of acute lung injury based on network pharmacology and differentially expressed genes of macrophages,and verify by experiments.Methods Cytoscape software was used to construct an compound-target-pathway network through PharmMapper,GeneCards and OMIM databases.The protein interaction network was established with the STRING database,and the core targets were speculated.GO and KEGG enrichment analysis were conducted on the targets with Metascape database.The macrophage chip data were downloaded,and the differential genes were obtained by GEO2R.KEGG signaling pathway analysis of differential genes were performed by DAVID platform.The expression of M1-polarized and M2-polarized macrophage F4/80,CD11b,CD86 and CD206 were detected by flow cytometry.PI3K and Akt protein expression in the lung were detected by Western blot.Results Ampelopsin had 102 targets intersection with acute lung injury,which were mainly enriched in PI3K/Akt,JAK/STAT,and Estrogen signaling pathway.Molecular docking showed that ampelopsin had good binding activity with MMP9 and IGF1.Differentially expressed genes were enriched in PI3K-Akt,cytokine-cytokine receptor interaction and chemokine signaling pathway.Flow cytometry showed that ampelopsin significantly increased the percentage of M2-polarized,reduced the proportion of M1/M2 and the expression of PI3K and Akt in lung.Conclusion Network pharmacology and animal experiments confirm that ampelopsin may alleviate ALI/ARDS by regulating macrophage polarization through PI3K/Akt pathway,which provides a theoretical basis to elucidate the mechanism of ampelopsin for ALI/ARDS.

关 键 词：双氢杨梅树皮素 急性肺损伤 网络药理学 巨噬细胞极化 

分 类 号：R285[医药卫生—中药学]