过表达IRG1抑制NLRP3炎症小体改善哮喘小鼠气道炎症  

Overexpression of IRG1 ameliorates airway inflammation of asthmatic mice by inhibiting NLRP3 inflammasome

作　　者：陈雪[1] 宁谦[1] CHEN Xue;NING Qian(Department of Respiratory and Critical Care Medicine,First Affiliated Hospital of Xi'an Jiaotong University Health Science Center,Xi'an 710061,China)

机构地区：[1]西安交通大学医学院第一附属医院呼吸与危重症医学科,西安710061

出　　处：《山西医科大学学报》2025年第2期134-141,共8页Journal of Shanxi Medical University

基　　金：陕西省重点研发计划项目(2022SF-141)。

摘　　要：目的 研究免疫响应基因1(IRG1)调控哮喘小鼠气道炎症的作用和机制。方法 将小鼠分为对照组、模型组、模型+AAV-Ctrl组和模型+AAV-IRG1组。构建表达IRG1重组腺相关病毒(AAV),采用气管插管术注射到小鼠肺部。采用卵清蛋白(OVA)诱导建立小鼠哮喘模型。采用Western blot检测小鼠肺组织中IRG1蛋白表达水平。采用液相色谱-质谱联用技术检测衣康酸在小鼠肺组织中的含量。采用苏木精-伊红染色检测小鼠肺组织病理。采用酶联免疫吸附试验检测小鼠支气管肺泡灌洗液(BALF)中炎症因子白细胞介素-4(interleukin-4,IL-4)、IL-5和IL-13的水平。采用全自动细胞分析仪检测小鼠BALF中炎症细胞数量。采用Western blot检测小鼠肺组织中核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、含CARD结构域的凋亡相关颗粒样蛋白(ASC)和半胱氨酸蛋白酶-1(Caspase-1)的蛋白表达水平。采用ELISA检测小鼠肺组织中IL-1β和IL-18的水平。结果 与对照组比较,模型组和模型+AAV-Ctrl组IRG1表达水平和衣康酸含量显著升高(P<0.05);与模型组和模型+AAV-Ctrl组比较,模型+AAV-IRG1组IRG1表达水平和衣康酸含量显著增加(P<0.01)。与对照组比较,模型组和模型+AAVCtrl组气管壁增厚、管腔变窄、气管周围存在大量炎症细胞浸润;与模型组和模型+AAV-Ctrl组比较,模型+AAV-IRG1组气管壁厚度减少、管腔增大、气管周围炎症细胞浸润减轻。与对照组比较,模型组和模型+AAV-Ctrl组BALF中IL-4、IL-5和IL-13的表达水平显著增加(P<0.01),炎症细胞包括嗜酸性粒细胞、中性粒细胞和淋巴细胞的数量都显著增多(P<0.01);与模型组和模型+AAV-Ctrl组比较,模型+AAV-IRG1组BALF中炎症因子水平和炎症细胞数量都显著降低(P<0.01)。与对照组比较,模型组和模型+AAV-Ctrl组NLRP3、ASC、Caspase-1、IL-1β和IL-18的表达水平显著升高(P<0.01);与模型组和模型+AAV-Ctrl组比价,模型+AAV-IRG1组NLRP3、ASObjective To investigate the role and mechanism of immune responsive gene 1(IRG1)in regulating airway inflammation of asthmatic mice.Methods The mice were divided into control group,model group,model+AAV-Ctrl group,and model+AAV-IRG1 group.The recombinant adeno-associated virus(AAV)expressing IRG1 was constructed and injected into the lungs of mice using tra-cheal intubation.The mouse model of asthma was established by ovalbumin(OVA)induction.The expression level of IRG1 protein in lung tissues was measured by Western blot.The content of itaconate in lung tissues was detected by liquid chromatography-mass spec-trometry.The pathological changes in mouse lung tissues were examined by hematoxylin and eosin(HE)staining.The levels of inflam-matory cytokines including interleukin-4(IL-4),IL-5,and IL-13 in the bronchoalveolar lavage fluid(BALF)were detected by enzyme linked immunosorbent assay(ELISA).The number of inflammatory cells in BALF was counted with an automatic cell analyzer.The pro-tein expression levels of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),and Caspase-1 in lung tissues were assessed by Western blot.The levels of IL-1βand IL-18 in lung tissues were measured using ELISA.Results Compared with control group,IRG1 expression level and itaconate content were sig-nificantly increased in model group and model+AAV-Ctrl group(P<0.05).Compared with model group and model+AAV-Ctrl group,IRG1 expression level and itaconate content were significantly increased in model+AAV-IRG1 group(P<0.01).Compared with con-trol group,the mice in model group and model+AAV-Ctrl group exhibited thickened tracheal walls,narrowed lumens,and increased infiltration of inflammatory cells around the trachea.Compared with model group and model+AAV-Ctrl group,the tracheal walls of mice were thinned in model+AAV-IRG1 group,lumens were enlarged,and the infiltration of inflammatory cells around the trachea was reduced.Compared with control group,the l

关 键 词：哮喘 气道炎症 肺组织损伤 IRG1 NLRP3炎症小体 腺相关病毒 衣康酸 

分 类 号：R562.2[医药卫生—呼吸系统]