抑制肝激酶B1上调M1肺泡巨噬细胞极化促进过敏性哮喘气道炎症反应  

Inhibition of LKB1 upregulation suppresses M1 alveolar macrophage polarization and promotes allergic asthmatic airway inflammatory responses

作　　者：兰舰 张睿昱 杜慧 刘智 雷雨溪 赵东赤[1] LAN Jian;ZHANG Ruiyu;DU Hui;LIU Zhi;LEI Yuxi;ZHAO Dongchi(Dept.of Pediatrics,Zhongnan Hospital of Wuhan University,Wuhan 430071,Hubei,China)

机构地区：[1]武汉大学中南医院儿科,湖北武汉430071

出　　处：《武汉大学学报(医学版)》2025年第3期291-297,385,共8页Medical Journal of Wuhan University

摘　　要：目的:探究肝激酶B1(LKB1)是否通过影响肺泡巨噬细胞(AMs)极化进而影响过敏性哮喘气道炎症。方法:将20只雌性BALB/c小鼠随机分为对照组、哮喘组、siRNA(si-LKB1)干预组和错配siRNA对照组4组,每组5只。使用卵清蛋白(OVA)致敏激发,建立急性过敏性哮喘的小鼠模型。在激发前1 d通过小鼠尾静脉注射分别给予si-LKB1和错配siRNA,并在连续激发3 d后,取肺泡灌洗液和肺组织检测。蛋白免疫印迹法检测肺组织LKB1表达水平;qPCR检测肺组织中mRNA水平,验证siRNA的敲低效果;HE和PAS染色检测小鼠肺组织炎症损伤;流式细胞术(FCS)检测肺泡灌洗液中细胞因子水平和AMs极化表型。结果:过敏性哮喘小鼠肺组织中LKB1表达比对照组明显增加(约为1.84倍)。抑制LKB1后,HE和PAS染色显示小鼠气道炎性细胞浸润加重,气管壁增厚更明显,杯状细胞明显增生。相较于哮喘组,敲低LKB1后肺泡灌洗液中TNF-α[(298.36±22.18)pg·mL^(-1) vs(218.78±35.83)pg·mL^(-1)]、IL-4[(26.70±0.87)pg·mL^(-1) vs(21.45±0.82)pg·mL^(-1)]升高;而IL-10[(9.02±1.27)pg·mL^(-1) vs(19.26±0.75)pg·mL^(-1)]降低;灌洗液中AMs占比增加[(18.57±0.73)%vs(7.49±0.81)%],且AMs中M1型巨噬细胞占比显著增加[(63.40±1.49)%vs(23.16±2.10)%]。哮喘组肺组织中LKB1蛋白表达水平增加[正常对照组(1.00±0.11)vs哮喘组(1.84±0.07),P<0.05]。结论:LKB1可以抑制AMs向M1方向极化并且减轻过敏性哮喘气道炎症的作用。抑制LKB1表达可以上调气道中TNF-α、IL-4等促炎因子的表达,促进气道中AMs聚集并向M1方向极化,并加重过敏性哮喘气道炎症。Objective:To investigate whether liver kinase B1(LKB1)influences airway inflammation in allergic asthma by affecting the polarization of alveolar macrophages(AMs).Methods:Twenty female BALB/c mice were randomly divided into four groups:control group,asthma group,siRNA(si-LKB1)intervention group,and mismatched siRNA control group,with five mice in each group.The mouse model of acute allergic asthma was established by sensitization with ovalbumin(OVA).One day before sensitization,si-LKB1 and mismatched siRNA were injected into the mice via the tail vein,respectively.After continuous stimulation for 3 days,alveolar lavage fluid and lung tissue were collected for detection.Western Blot was used to detect the expression level of LKB1 in mouse lung tissue;qPCR was used to detect the mRNA level in mouse lung tissue to verify the knockdown effect of siRNA;HE and PAS staining were performed to evaluate inflammation and injury in mouse lung tissues;Flow cytometry was used to detect the levels of cytokines and AMs polarization phenotype in bronchoalveolar lavage fluid.Results:In the lung tissues of mice with allergic asthma,the expression of LKB1 significantly increased by approximately 1.84 times as compared with the control group.Inhibiting LKB1 led to increased inflammatory cell infiltration,more pronounced thickening of the bronchial walls,and noticeable hyperplasia of goblet cells,as observed through HE and PAS staining.Compared with those in the asthma group,knocking down LKB1 resulted in elevated levels of TNF-α((298.36±22.18)pg·mL^(-1) vs(218.78±35.83)pg·mL^(-1))and IL-4((26.70±0.87)pg·mL^(-1) vs(21.45±0.82)pg·mL^(-1))in alveolar lavage fluid;meanwhile,IL-10((9.02±1.27)pg·mL^(-1) vs(19.26±0.75)pg·mL^(-1))decreased.The proportion of AMs the alveolar lavage fluid increased as(18.57±0.73)%vs(7.49±0.81%),and the proportion of M1 type macrophages to AMs increased as(63.40±1.49)%vs(23.16±2.10)%.Conclusion:LKB1 can inhibit the polarization of AMs towards M1 and alleviate allergic asthma airway inflammation

关 键 词：过敏性哮喘 LKB1 巨噬细胞极化 气道炎症 

分 类 号：R725.6[医药卫生—儿科]