基于生物信息学分析的变应性鼻炎小鼠模型中的枢纽基因鉴定

Hub Genes Identification in a Murine Model of Allergic Rhinitis Based on Bioinformatics Analysis.

作者信息

Chen Le, Shi Le, Ma Yue, Zheng Chunquan

机构信息

Department of Otolaryngology-Head and Neck Surgery, Eye, Ear, Nose, and Throat Hospital, Fudan University, Shanghai, China.

Shanghai Key Clinical Disciplines of Otorhinolaryngology, Shanghai, China.

出版信息

Front Genet. 2020 Aug 25;11:970. doi: 10.3389/fgene.2020.00970. eCollection 2020.

Abstract

This study aimed to identify allergic rhinitis (AR)-related hub genes and functionally enriched pathways in a murine model. Dataset GSE52804 (including three normal controls and three AR mice) was downloaded from Gene Expression Omnibus (GEO). Differentially expressed genes (DEGs) were identified. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and protein-protein interaction (PPI) analyses of DEGs were performed to identify the hub genes in AR. The DEGs were classified into different modules by using the weighted gene co-expression network analysis (WGCNA). Moreover, to verify the potential hub genes, nasal mucosa tissues were obtained from murine AR models ( = 5) and controls ( = 5), and qRT-PCR and Western blot were performed. In this study, a total of 634 DEGs were identified. They were significantly enriched in 14 GO terms, such as integral component of membrane, plasma membrane, and G-protein-coupled receptor signaling pathway. Meanwhile, there were eight terms of KEGG pathways significantly enriched, such as Olfactory transduction, Cytokine-cytokine receptor interaction, and TNF signaling pathway. The top 10 hub genes (Rtp1, Rps27a, Penk, Cxcl2, Gng8, Gng3, Cxcl1, Cxcr2, Ccl9, and Anxa1) were identified by the PPI network. DEGs were classified into seven modules by WGCNA. According to qRT-PCR validation of the five genes of interest (Rtp1, Rps27a, Penk, Cxcl2, and Anxa1), the expression level of Rtp1 mRNA was significantly decreased in the AR group compared with the control group, while there are enhanced Rps27a, Penk, Cxcl2, and Anxa1 mRNA expressions in the AR mice group compared with the control group. Western blot was also performed to further explore the expression of Anxa1 in the protein level, and the results showed a similar expression trend.

摘要

本研究旨在鉴定小鼠模型中与变应性鼻炎(AR)相关的枢纽基因及功能富集通路。数据集GSE52804(包括三只正常对照小鼠和三只AR小鼠)从基因表达综合数据库(GEO)下载。鉴定差异表达基因(DEG)。对DEG进行基因本体论(GO)、京都基因与基因组百科全书(KEGG)通路及蛋白质-蛋白质相互作用(PPI)分析,以鉴定AR中的枢纽基因。通过加权基因共表达网络分析(WGCNA)将DEG分类到不同模块中。此外,为验证潜在的枢纽基因,从小鼠AR模型(n = 5)和对照组(n = 5)获取鼻黏膜组织,并进行qRT-PCR和蛋白质印迹分析。本研究共鉴定出634个DEG。它们在14个GO术语中显著富集,如膜的整合成分、质膜和G蛋白偶联受体信号通路。同时,有8个KEGG通路术语显著富集,如嗅觉转导、细胞因子-细胞因子受体相互作用和TNF信号通路。通过PPI网络鉴定出前10个枢纽基因(Rtp1、Rps27a、Penk、Cxcl2、Gng8、Gng3、Cxcl1、Cxcr2、Ccl9和Anxa1)。通过WGCNA将DEG分类到7个模块中。根据对五个感兴趣基因(Rtp1、Rps27a、Penk、Cxcl2和Anxa1)的qRT-PCR验证,与对照组相比,AR组中Rtp1 mRNA的表达水平显著降低,而与对照组相比,AR小鼠组中Rps27a、Penk、Cxcl2和Anxa1 mRNA的表达增强。还进行了蛋白质印迹分析以进一步探究Anxa1在蛋白质水平的表达,结果显示出相似的表达趋势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f4c/7477359/74a04c53c19f/fgene-11-00970-g001.jpg

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