慢性阻塞性肺疾病(COPD)模型大鼠肺组织中NLRP3炎症小体表达水平的变化及意义  

Changes and Significance of NLRP3 Inflammasome in the Rats Model of Chronic Obstructive Pulmonary Disease(COPD)

作　　者：杨文林[1] 顾慧玲[1] 倪红燕[1] 王海峰[1] YANG Wen- lin,GU Hui -ling,NI Hong -yan,WANG Hai -feng(Department of Respiratory, Baoshan Branch of Shanghai First People＇s Hospital, Shanghai 200940, China)

机构地区：[1]上海市第一人民医院宝山分院呼吸科,上海200940

出　　处：《现代检验医学杂志》2018年第6期69-72,共4页Journal of Modern Laboratory Medicine

基　　金：上海市宝山区科学技术委员会(编号:14-E-12)

摘　　要：目的探讨NOD样受体热蛋白结构相关蛋白3(NLRP3)炎症小体在慢性阻塞性肺疾病(COPD)模型中的变化及意义。方法将30只雄性大鼠随机分为2组,正常对照组(n=10只)给予自由饮水和摄食,喂养75天;吸烟组(n=20只)采用单纯吸入香烟烟雾的方法制造大鼠COPD模型,连续75天后再根据大鼠最大呼气流速(PEF)及气道肺组织病理变化分为吸烟COPD组(n=9只)和吸烟非COPD组(n=11只)。收集支气管肺泡灌洗液(BALF)计算巨噬细胞(AMC)、中性粒细胞(NEU)和淋巴细胞(LYM)的绝对计数。采用酶联免疫吸附试验(ELISA)、逆转录-聚合酶链反应(RT-PCR)技术分别测定BALF,肺组织中NLRP3炎症小体的浓度以及NLRP3mRNA的相对表达量。结果与正常对照组比较,吸烟非COPD组、吸烟COPD组BALF中NEU(×106/L)(524.2±73.1,916.9±84.8vs 106.6±31.8),AMC(×106/L)(1 570.9±273.5,2 307.8±410.4vs 496.6±61.7)和LYM(×106/L)(72.1±14.7,115.4±23.8vs 21.6±4.2),计数明显升高,吸烟COPD组BALF中NEU,AMC和LYM计数亦较吸烟非COPD组明显升高(F=350.59,100.57,82.63,均P<0.05)。与正常对照组比较,吸烟非COPD组、吸烟COPD组BALF(pg/ml)(107.8±23.5,126.7±34.9vs 76.7±15.1),肺组织(pg/ml)(118.8±33.3,147.4±40.2vs 84.1±21.5)中NLRP3炎症小体的浓度明显升高,与吸烟非COPD组比较,吸烟COPD组BALF,肺组织中NLRP3炎症小体的浓度亦明显升高(F=9.53,9.17,均P<0.05)。吸烟非COPD组和吸烟COPD组肺组织NLRP3mRNA相对表达量均显著高于正常对照组(P<0.05),吸烟COPD组肺组织NLRP3mRNA相对表达量显著高于吸烟非COPD组(P<0.05)。BALF,肺组织中NLRP3炎症小体浓度、肺组织NLRP3mRNA相对表达量与NEU,AMC,LYM计数均呈显著正相关(P<0.05)。结论 NLRP3炎症小体参与了COPD慢性炎症的发生、发展,与肺组织炎症细胞以及气道病理改变密切相关,阻断NLRP3炎性小体的活化,有望成为治疗COPD新的靶点和方向。Objective To explore the changes and significance of NLRP3 inflammasome in the rats of chronic obstructive pulmonary disease(COPD).Methods 30 male rats were divided randomly into two groups.Control group(n=10)was given with drink and eat freely for 75 days.Smoking group(n=20)was given with inhaling cigarette smoke to establish rat model of COPD.After 75 days,the rats in smoking group were divided into smoking COPD group(n=9)and smoking non-COPD group(n=11)according to the maximum expiratory velocity(PEF)and pathologic changes of airway lung tissue.Bronchoalveolar lavage fluid(BALF)was collected to calculate the count of macrophages(AMC),neutrophils(NEU),and lymphocyte(LYM).Enzyme-linked immunosorbent(ELISA),reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the NLRP3 inflammasome in BALF and lung tissue,and the relative expression of NLRP3 mRNA.ResultsCompared with control group,the count of NEU(×106/L)(524.2±73.1,916.9±84.8 vs 106.6±31.8),AMC(×106/L)(1 570.9±273.5,2 307.8±410.4 vs 496.6±61.7)and LYM(×106/L)(72.1±14.7,115.4±23.8 vs 21.6±4.2)in BALF of smoking COPD group and smoking non-COPD group were increased significantly,these indicators were also increased significantly in smoking COPD group compared with smoking non-COPD groups(F=350.59,100.57 and 82.63,all P<0.05).Compared with control group,the count of NEU,AMC,LYM in BALF of smoking COPD group and smoking non-COPD group were increased significantly,these indicators were also increased significantly in smoking COPD group compared with smoking non-COPD groups(P<0.05).Compared with control group,the concentration of NLRP3 inflammasome in BALF(pg/ml)(107.8±23.5,126.7±34.9 vs 76.7±15.1),lung tissue(118.8±33.3 pg/ml,147.4±40.2 pg/ml vs 84.1±21.5 pg/ml)of smoking COPD group and smoking non-COPD group was increased significantly,and these indicators were also increased significantly in smoking COPD group compared with smoking non-COPD groups(F=9.53,9.17,all P<0.05).The NLRP3 mRNA relative expression in lung tissue of smok

关 键 词：NOD样受体热蛋白结构域相关蛋白3炎症小体 慢性阻塞性肺疾病 大鼠COPD模型 

分 类 号：R563[医药卫生—呼吸系统] R-332[医药卫生—内科学]