大气细颗粒物通过TLR4/NF-κB通路诱导鼻黏膜上皮细胞炎症反应  

PM_(2.5)induces inflammatory response in nasal epithelial cells through TLR4/NF-κB pathway

作　　者：杨晓喆 赵妍 青卉 王向东[1,2,3] 张罗[1,2,3] YANG Xiaozhe;ZHAO Yan;QING Hui;WANG Xiangdong;ZHANG Luo(Department of Otolaryngology Head and Neck Surgery,Beijing Tongren Hospital,Capital Medical University,Key Laboratory of Otolaryngology Head and Neck Surgery(Capital Medical University),Ministry of Education,Beijing 100730,China;Beijing Institute of Otolaryngology,Beijing Laboratory of Allergic Diseases,Beijing Municipal Education Commission,Beijing Key Laboratory of Nasal Diseases,Beijing 100005,China;Department of Allergy,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China)

机构地区：[1]首都医科大学附属北京同仁医院耳鼻咽喉头颈外科,耳鼻咽喉头颈科学教育部重点实验室(首都医科大学),北京100730 [2]北京市耳鼻咽喉科研究所,过敏性疾病北京实验室,鼻病研究北京市重点实验室,北京100005 [3]首都医科大学附属北京同仁医院变态反应科,北京100730

出　　处：《山东大学学报（医学版）》2023年第9期47-56,共10页Journal of Shandong University:Health Sciences

基　　金：国家重点研发计划(2022YFC2504100);教育部长江学者与创新团队发展计划(IRT13082);中国医学科学院医学与健康科技创新工程项目(2019-I2M-5-022);首都临床特色诊疗技术研究及转化应用重点项目(Z211100002921057);首都卫生发展科研专项重点攻关项目(2022-1-1091);国家自然科学基金(82101190,82171110,82000962,81970852);北京市自然科学基金(7222024);北京市卫生系统高层次公共卫生人才建设项目领军人才(领军人才-01-08,领军人才-02-09);北京市医管中心“青苗”人才计划(QML20230201)。

摘　　要：目的探讨阐明TLR4/NF-κB信号通路在PM_(2.5)诱导鼻黏膜上皮细胞炎症反应中的作用机制。方法参考健康人鼻黏膜上皮细胞系(HNEpCs)构建PM_(2.5)暴露模型。暴露后提取RNA利用Illumina测序仪进行转录组分析。荧光定量PCR(qRT-PCR)分析TLR4、NFKB1、NFKB1A基因的RNA表达水平。结果PM_(2.5)可引起HNEpCs毒性及转录组异常调控,导致TLR4、NFKBIA及IL1B基因的异常表达,KEGG通路分析发现所参与的TLR4/NF-κB通路显著富集。qRT-PCR验证实验发现,经过PM_(2.5)暴露后TLR4基因上调,NFKB1基因下调,与此同时抑制NF-κB的NFKB1A(NF-κB抑制因子α)显著下调,故而在一定程度上导致TLR4/NF-κB通路异常调控。结论本研究发现HNEpCs在PM_(2.5)暴露后,TLR4/NF-κB通路异常调控,可能在PM_(2.5)暴露所诱发的鼻黏膜炎症反应中起到关键作用。Objective To investigate the mechanism of TLR4/NF-κB signaling pathway in the inflammatory response of nasal mucosal epithelial cells induced by fine particulate matter(PM_(2.5)).Methods After healthy human nasal epithelial cells(HNEpCs)were exposed to PM_(2.5),the RNA was extracted for transcriptome analysis by Illumina sequencer.The expressions of TLR4,NFKB1 and NFKB1A were detected with quantitative real-time PCR(qRT-PCR).Results PM_(2.5)caused cell toxicity and abnormal regulation of transcriptome of HNEpCs,and abnormal expressions of TLR4,NFKBIA and IL1B.KEGG pathway analysis indicated that TLR4/NF-κB pathway was significantly enriched.The qRT-PCR validation experiment revealed that after PM_(2.5)exposure,TLR4 was up-regulated while NFKB1 was down-regulated,and NFKBIA,which inhibited NF-κB,was significantly down-regulated,thus leading to abnormal regulation of the TLR4/NF-κB pathway.Conclusion The abnormal regulation of TLR4/NF-κB in HNEpCs may play a crucial role in the nasal mucosa inflammatory response induced by PM_(2.5)exposure.

关 键 词：鼻黏膜上皮细胞系 大气细颗粒物 暴露模型 基因异常表达 荧光定量分析 转录组分析 鼻黏膜炎症 

分 类 号：R765.25[医药卫生—耳鼻咽喉科]