Sun L W, Liu Z Y, Sha J C, Meng C D, Zhu D D
Department of Otolaryngology Head and Neck Surgery, China-Japan Union Hospital of Jilin University, Changchun 130033, China Jilin Provincial Key Laboratory of Precise Diagnosis and Treatment of Upper Airway Allergic Diseases, Changchun 130033, China.
School of Life Science, Jilin University, Changchun 130012, China.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2022 Apr 7;57(4):425-432. doi: 10.3760/cma.j.cn115330-20210630-00397.
Transcriptome sequencing and bioinformatics analysis were performed on the gene expression of nasal epithelial cells in patients with seasonal allergic rhinitis (AR) and perennial AR, so as to obtain the differences in the gene expression of nasal epithelial cells between seasonal AR and perennial AR. The human nasal epithelial cell line(HNEpC) was cultured , treated with 100 μg/ml mugwort or house dust mite (HDM) extracts for 24 hours. Total cell RNA was extracted, and quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression of cytokines, including IL-6, IL-8, IL-33 and thymic stromal lymphopoietin (TSLP). From November 2019 to November 2020, 3 seasonal AR patients, 3 perennial AR patients, and 3 healthy controls who attended the Department of Otolaryngology Head and Neck Surgery, China-Japan Union Hospital of Jilin University were analyzed. The patients' primary nasal epithelial cells were cultured , treated with corresponding allergens for 24 hours. Total RNA was extracted for transcriptome sequencing, and the sequencing results were analyzed by bioinformatics. The qPCR results showed that the cytokines IL-6, IL-8, IL-33 and TSLP of HNEpC treated with mugworts extracts and HDM extracts had the same trend of change. After the nasal epithelial cells from patients with seasonal AR and perennial AR were treated with corresponding allergens, there were differences in biological processes and signal pathways between those and control. Gene ontology (GO) enrichment analysis showed that the differentially expressed genes (DEG) in AR patients allergic to mugwort were mainly enriched in the oxidation-reduction process, the negative regulation of apoptosis process, and the cell adhesion; the DEG in AR patients allergic to HDM were mainly enriched in cell adhesion, the negative regulation of cell proliferation and the response to drug. Enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway showed that the DEG of AR patients allergic to mugwort were significantly enriched in arachidonic acid metabolism, p53 signaling pathway and transforming growth factor β (TGF-β) signaling pathway, while the DEG of AR patients allergic to HDM were mainly enriched in cells cycle, Fanconi anemia pathway and DNA replication. Gene Set Enrichment Analysis (GSEA) showed that the inflammatory response, TNF-α/NF-κB signaling pathway and IL-2/STAT5 signaling pathway were significantly up-regulated in AR patients allergic to mugwort, indicating the promotion of inflammatory response; and AR patients allergic to HDM had significant down-regulation of G2M, E2F, and MYC, indicating the inhibition of cell proliferation. The protein-protein interaction network showed that TNF and CDK1 were the most interacting proteins in mugwort and HDM allergic AR patients, respectively. Seasonal AR and perennial AR may affect the different biological processes and signal pathways of nasal epithelial cells, leading to differences in the occurrence and development of AR.
对季节性变应性鼻炎(AR)患者和常年性AR患者的鼻上皮细胞基因表达进行转录组测序和生物信息学分析,以获得季节性AR和常年性AR之间鼻上皮细胞基因表达的差异。培养人鼻上皮细胞系(HNEpC),用100μg/ml艾蒿或屋尘螨(HDM)提取物处理24小时。提取总细胞RNA,采用定量实时聚合酶链反应(qPCR)检测细胞因子IL-6、IL-8、IL-33和胸腺基质淋巴细胞生成素(TSLP)的表达。2019年11月至2020年11月,对吉林大学中日联谊医院耳鼻咽喉头颈外科就诊的3例季节性AR患者、3例常年性AR患者和3例健康对照进行分析。培养患者的原代鼻上皮细胞,用相应变应原处理24小时。提取总RNA进行转录组测序,并通过生物信息学分析测序结果。qPCR结果显示,用艾蒿提取物和HDM提取物处理的HNEpC的细胞因子IL-6、IL-8、IL-33和TSLP具有相同的变化趋势。季节性AR和常年性AR患者的鼻上皮细胞用相应变应原处理后,与对照组相比,其生物学过程和信号通路存在差异。基因本体(GO)富集分析显示,对艾蒿过敏的AR患者中差异表达基因(DEG)主要富集于氧化还原过程、凋亡过程的负调控和细胞黏附;对HDM过敏的AR患者中DEG主要富集于细胞黏附、细胞增殖的负调控和药物反应。京都基因与基因组百科全书(KEGG)信号通路富集分析显示,对艾蒿过敏的AR患者的DEG在花生四烯酸代谢、p53信号通路和转化生长因子β(TGF-β)信号通路中显著富集,而对HDM过敏的AR患者的DEG主要富集于细胞周期、范可尼贫血通路和DNA复制。基因集富集分析(GSEA)显示,对艾蒿过敏的AR患者中炎症反应、TNF-α/NF-κB信号通路和IL-2/STAT5信号通路显著上调,表明炎症反应增强;对HDM过敏的AR患者中G2M、E2F和MYC显著下调,表明细胞增殖受到抑制。蛋白质-蛋白质相互作用网络显示,TNF和CDK1分别是艾蒿和HDM过敏AR患者中相互作用最多的蛋白质。季节性AR和常年性AR可能影响鼻上皮细胞的不同生物学过程和信号通路,导致AR发生发展的差异。