基于网络药理学的小儿定喘颗粒抗呼吸道合胞病毒肺炎的作用机制研究

Study on the mechanism of Xiaoer Dingchuan Cranules against respiratory syncytial virus pneumonia based on network pharmacology

关键词:  网络药理学  小儿定喘颗粒  呼吸道合胞病毒  活性成分  靶点  作用机制

Key words:  network pharmacology  Xiaoer Dingchuan Granule  RSV  active ingredients  target  mechanism of action

摘要:

[目的] 本研究初步探究了小儿定喘颗粒抗呼吸道合胞病毒的药理作用，并通过网络药理学及生物信息学技术探讨小儿定喘颗粒治疗呼吸道合胞病毒肺炎（RSV）的作用机制。[方法] 通过建立RSV感染的A549细胞模型以考察小儿定喘颗粒抗RSV的治疗效果；利用中药系统药理学数据库（TCMSP）获取小儿定喘颗粒的主要化学成分及作用靶点，利用Uniprot数据库对获取的作用靶点进行规范化处理；通过GeneCards及OMIM数据库获取RSV相关靶点；利用Venny 2.1.0绘图网站构建Venn图，得到小儿定喘颗粒与RSV的交集靶点；应用String平台构建蛋白质-蛋白质相互作用（PPI）网络；通过David数据库对交集靶点进行基因本体论（GO）及京都基因与基因组百科全书（KEGG）通路富集分析，使用微生信网站绘制结果气泡图，相关结果采用Cytoscape 3.9.0软件进行可视化及网络拓扑结构分析。使用分子对接及酶联免疫吸附实验对上述预测核心靶点进行验证。[结果] 体外细胞实验表明小儿定喘颗粒对RSV诱导的细胞病变具有抑制作用，经过数据整理及分析共筛选出小儿定喘颗粒的主要化学成分194个，潜在作用靶点282个，其中涉及RSV靶点135个。“中药-成分-疾病靶点”网络显示，槲皮素、山柰酚、异鼠李素等是小儿定喘颗粒治疗RSV的主要活性成分，肿瘤坏死因子（TNF）、白蛋白（ALB）、白细胞介素-6（IL-6）、RAC-α-丝氨酸/苏氨酸蛋白激酶（AKT1）、白细胞介素-1β（IL-1β）等是小儿定喘颗粒治疗RSV的核心靶点。通过GO功能富集分析得到822条生物进程条目，138条分子功能条目，89条细胞成分条目，KEGG富集分析发现核心靶点主要作用于TNF、IL-17、磷脂酰肌醇3激酶（PI3K）-蛋白激酶B（Akt）等信号通路，分子对接、酶联免疫吸附实验进一步验证了上述结果。[结论] 初步揭示了小儿定喘颗粒可能通过多成分、多靶点和多通路发挥治疗RSV的作用，为拓宽其临床应用提供理论基础。

Abstract:

[Objective] In this study，the pharmacological effects of Xiaoer Dingchuan Cranules against respiratory syncytial virus were preliminarily investigated，and the mechanism of action of Xiaoer Dingchuan Cranules in the treatment of respiratory syncytial virus（RSV） was explored by network pharmacology and bioinformatics technology. [Methods] The A549 cell model of RSV infection was established to investigate the therapeutic effect of Xiaoer Dingchuan Granules against RSV；The main chemical components and targets of Xiaoer Dingchuan Granules were obtained by using the Pharmacological Database of Traditional Chinese Medicine System（TCMSP），and the obtained targets were standardized by Uniprot database. Obtain RSV-related targets through the GeneCards and OMIM databases. The Venny 2.1.0 mapping website was used to construct the Venn map to obtain the intersection target of Xiaoer Dingchuan Granules and RSV. Apply the String platform to build a protein-protein interaction（PPI） network. The intersection targets were analyzed through the David database for gene ontology（GO） and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment，and the results were mapped using the microbiotic website，and the relevant results were visualized and analyzed by Cytoscape 3.9.0 software. The above predicted core targets were verified using molecular docking and enzyme-linked immunosorbent assays. [Results] In vitro cell experiments showed that Xiaoer Dingchuan Granules had inhibitory effect on RSV-induced cytopathies，a total of 194 main chemical components and 282 potential targets were screened for Xiaoer Dingchuan Granules through data collection and analysis，including 135 RSV targets. The “traditional Chinese medicine-ingredients-disease target” network shows that quercetin，kaempferol，isorhamnetine，etc are the main active ingredients in the treatment of RSV in Xiaoer Dingchuan Granules，tumor necrosis factor（TNF），albumin（ALB），interleukin-6（IL-6），RAC-α-serine/threonine protein kinase（RAC-alpha serine/threonine-protein kinase，AKT1），interleukin-1β（IL-1β） and others are the core targets of Xiaoer Dingchuan Granules for the treatment of RSV. Through GO function enrichment analysis，822 biological process entries，138 molecular function items and 89 cell component entries were obtained，KEGG enrichment analysis found that the core targets mainly acted on TNF，IL-17，PI3K-Akt and other signaling pathways，molecular docking and enzyme-linked immunosorbent assays further verified the above results. [Conclusion] It was preliminarily revealed that Xiaoer Dingchuan Granules may play a role in the treatment of RSV through multi-component，multi-target and multi-pathway，which provided a theoretical basis for broadening its clinical application.