DEC2缺失促进感染后咳嗽高发及盐酸麻黄碱的治疗作用机制  

DEC2 deficiency promotes high incidence of post infectious cough and the therapeutic mechanism of ephedrine hydrochloride

作　　者：张庆国 杨雯越 王悦 张玥 姚成芳 ZHANG Qingguo;YANG Wenyue;WANG Yue;ZHANG Yue;YAO Chengfang(School of Clinical and Basic Medicine,Shandong First Medical University,Shandong Academy of Medical Sciences,Jinan 250117,China)

机构地区：[1]山东第一医科大学,山东省医学科学院临床与基础医学院,济南250117

出　　处：《中国免疫学杂志》2025年第4期854-860,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金面上项目(82074088);山东省重点研发计划重大科技创新工程项目(2022CXGC020514)。

摘　　要：目的:探讨DEC2缺失促进感染后咳嗽(PIC)高发及盐酸麻黄碱(EH)的治疗作用机制。方法:利用C57BL/6野生型(WT)小鼠和DEC2敲除(DEC2-KO)小鼠,采用脂多糖(LPS)滴鼻加烟雾刺激等建立PIC模型,部分模型小鼠采用EH干预。在造模第12、14、16、17天计数辣椒素激发的咳嗽;利用单细胞测序、流式细胞术、RT-qPCR等技术检测PIC造模及EH干预前后小鼠肺脏局部IL-1、IL-6、IFN-γ等炎症因子及受体的表达;利用小鼠肺脏CD45−原代细胞培养体系,在LPS刺激或EH干预下,采用RT-qPCR分析炎症因子及其受体,以及NF-κB信号分子的表达。结果:WT-PIC模型小鼠DEC2表达显著升高,而DEC2缺失则导致DEC2-KO-PIC模型小鼠咳嗽频率急剧升高,为WT-PIC小鼠的2倍;DEC2缺失加剧了肺脏LPS诱导的炎症反应,特别是肺脏CD45−细胞表达IL-1、IL-6、γ-干扰素受体(IFN-γR)、IL-1Ra、IL-6Ra等炎症因子及其受体的能力显著升高(P<0.01),NF-κB信号过度激活(P<0.01);EH干预后可以显著下调DEC2-KO-PIC模型小鼠的咳嗽频率(P<0.001),抑制肺脏IL-1、IL-6、IFN-γ等炎症因子的分泌,降低肺脏CD45−细胞IL-1Ra、IL-6Ra、IFN-γR等炎症因子受体的表达(P<0.01),下调DEC2缺失诱导的NF-κB信号的表达和活化(P<0.01)。结论:DEC2缺失可以激活NF-κB信号,促进肺脏CD45−细胞炎症因子及其受体高表达,是DEC2缺失后PIC高发的关键因素,EH能抑制DEC2缺失导致的NF-κB信号过度活化和肺脏局部基质细胞的炎症特征,是有效治疗PIC的关键药理机制。Objective:To explore the mechanism of DEC2 deficiency promoting high incidence of post infectious cough(PIC)and the therapeutic effects of ephedrine hydrochloride(EH).Methods:C57BL/6 wild-type(WT)mice and DEC2-knockout(DEC2-KO)mice were treated with lipopolysaccharide(LPS)nose drops and smoke stimulation to establish PIC models,and some model mice were treated with EH intervention.Counting capsaicin-induced cough at 12,14,16 and 17 day of modeling;Single-cell sequencing,flow cytometry,RT-qPCR and other techniques were used to detect the expressions of IL-1,IL-6,IFN-γand other inflammatory factors and receptors in the lungs of mice before and after PIC modeling and EH intervention.The expressions of inflammatory factors and their receptors,as well as NF-κB signaling molecules,were analyzed by RT-qPCR using mouse lung CD45−primary cell culture system under LPS stimulation or EH intervention.Results:The transcription of DEC2 was increased in WT-PIC model mice,and the deficiency of DEC2 in DEC2-KO-PIC mice induced the cough frequency 2-fold higher than that in WT-PIC mice.The absence of DEC2 exacerbated LPS-induced pulmonary inflammatory response,especially increased expressions of IL-1,IL-6,IL-1Ra,IL-6Ra,IFN-γR in lung CD45−cells(P<0.01),and excessive activation of NF-κB signal(P<0.01).After EH treatment,the cough frequency in DEC2-KO-PIC mice was significantly reduced(P<0.001),the expression of IL-1,IL-6,IFN-γin lung and IL-1Ra,IL-6Ra,IFN-γR in pulmonary CD45−cells were down-regulated(P<0.01).The expression and activation of NF-κB induced by DEC2 deletion were significantly inhibited(P<0.01).Conclusion:The deficiency of DEC2 can activate NF-κB signal and promote the high expression of inflammatory cytokine and receptors in pulmonary CD45−cells,which is a risk factor in the high incidence of PIC.EH can inhibit the excessive-activation of NF-κB signal and the inflammatory characteristics of lung local stromal cells caused by DEC2 absence,and that is the main part of pharmacological mechanism for effe

关 键 词：DEC2 感染后咳嗽 肺脏炎症微环境 盐酸麻黄碱 

分 类 号：R392.5[医药卫生—免疫学]