基于网络药理学和分子对接的宣肺败毒方治疗急性呼吸窘迫综合征的机制及验证研究

Mechanism and confirmatory study of Xuanfei Baidu Decoction on treating ARDS based on network pharmacology and molecular docking

关键词:  宣肺败毒方  急性呼吸窘迫综合征  网络药理学  铁死亡

Key words:  Xuanfei Baidu Decoction  acute respiratory distress syndrome  network pharmacology  ferroptosis

摘要:

[目的] 采用网络药理学和分子对接技术探究宣肺败毒方治疗急性呼吸窘迫综合征（ARDS）的作用机制并进行验证。[方法] 通过中药系统药理学数据分析平台，对宣肺败毒方的组成药物麻黄、苍术、藿香、青蒿、虎杖、马鞭草、薏苡仁、芦根、葶苈子、苦杏仁、化橘红、甘草分别进行活性成分筛选，构建活性成分-靶点相互作用网络图，采用Genecards、SwissTargetPrediction和Uniprot数据库获取ARDS疾病靶点，筛选ARDS核心靶点，构建靶点间蛋白相互作用分析图，采用DAVID数据库对核心靶点进行GO和KEGG分析，采用Schrödinger软件对活性成分-核心靶点分子对接并计算结合能。通过脂多糖构建ARDS小鼠模型，造模1 d后经宣肺败毒方连续灌胃28 d，通过流式细胞及酶联免疫吸附（ELISA）实验分析肺泡灌洗液中白细胞介素（IL）-2、IL-4、IL-6、IL-10、肿瘤坏死因子（TNF）-α、干扰素（IFN）-γ、肺组织中谷胱甘肽（GSH）、丙二醛（MDA）、超氧化物歧化酶（SOD）和磷脂过氧化氢GSH过氧化物酶（GPX4）的变化，对网络药理学结果进行验证。[结果] 筛选得到宣肺败毒方治疗ARDS的活性成分205个，度值前8位的化合物分别是槲皮素、山柰酚、β-谷甾醇、豆甾醇、木犀草素、异鼠李素、柚皮苷和光甘草定。筛选得到核心靶点107个，度值排名前10位的核心靶点为Jun、IL-6、TP53、AKT1、ALB、VEGFA、STAT3、CASP3、IL-1β和PTGS2。生物信息学分析发现宣肺败毒方治疗ARDS与IL-7、TNF和Toll样受体等炎性反应相关信号通路有关，与PTGS2等铁死亡相关蛋白具有较低结合能。小鼠模型证实，宣肺败毒方可以显著降低脂多糖诱发的肺泡灌洗液中炎症因子异常升高和肺组织氧化还原失衡。[结论] 宣肺败毒方治疗ARDS的机制与缓解氧化损伤与细胞凋亡、调控炎症信号通路、减少炎症细胞募集浸润和细胞因子分泌有关，干预铁死亡途径可能是重要机制。

Abstract:

[Objective] Explored and confirmed the mechanism of Xuanfei Baidu Decoction on acute respiratory distress syndrome(ARDS) by network pharmacology and molecular docking. [Methods] The active components of mahuang,cangzhu,huoxiang,qinghao,huzhang,mabiancao,yiyiren,lugen,tinglizi,kuxingren,huajuhong and gancao in Xuanfei Baidu Decoction were collected and screened by TCMSP and active component-target network was constructed. Genecards,SwissTargetPrediction and Uniprot database was used to obtain the disease targets of ARDS and target proteins network was constructed,DAVID database was used to perform gene ontology(GO) and KEGG pathway enrichment analysis. Components and key targets were docked by Schrödinger software to verify the binding energy. In order to confirmed the result of network pharmacology,ARDS rat were constructed by lipopolysaccharides and continuously sponsored the stomach for 28 days by Xuanfei Baidu Decoction.The interleukin(IL)-2,IL-4,IL-6,IL-10 and tumor Necrsis Factor(TNF)-ɑ,Interfereon(IFN)-γ in alveolar lavage fluid,the Glutathione(GSH),Malondialdehyde(MDA),Superoxidismutase(SOD) and Glutathione Peroxidase 4(GPX4) in lung tissues were analyzed by flow cytometry and ELISA assay. [Results] There were 205 active components which quercetin,kaempferol, β-sitosterol,stigmasterol,luteolin,isorhamnetin,naringenin and glabridin were top 8 degree value components,107 Hub targets which Jun,IL-6,TP53,AKT1,ALB,VEGFA,STAT3,CASP3,IL-1β and PTGS2 were top 10 degree value targets in Xiaoxianxiong Decoction for ARDS treatment. Bioinformation analysis showed the signaling pathway of IL-7,TNF and Toll-like receptor was related to the treatment of ARDS by Xuanfei Baidu Decoction. There was a low binding energy between key active components and ferroptosis related protein such as PTGS2 by molecular docking. Xuanfei Baidu Decoction could revise the abnormal inflammatory factor in alveolar lavage fluid and oxidation-reduction disbalance in lung tissues which inducted by lipopolysaccharides in rat model. [Conclusion] The mechanism of Xuanfei Baidu Decoction treat ARDS was related with releasing the oxidative damage and apoptosis,regulating inflammation signaling pathway,reducing inflammatory recruitment and cytokines secretion,and ferroptosis inhibition may be play an important role.