NLRP3／IL-1β和IL-18在哮喘小鼠肺组织中的表达及与呼吸道炎症的关系  

Analysis of the roles of NLRP3 -dependent IL-1β and IL-18 in the development of allergic airway disease in mice with asthma

作　　者：梁海梅[1] 于化鹏[1] 夏虎[1] 邓火金[1] 樊慧珍[1] 龚雨新[1] 方泽葵[1] 郑燕妮[2] 刁建新[3] 

机构地区：[1]南方医科大学珠江医院呼吸内科,广州510282 [2]广东揭阳市人民医院呼吸内科 [3]南方医科大学中医药学院

出　　处：《中华微生物学和免疫学杂志》2013年第4期281-286,共6页Chinese Journal of Microbiology and Immunology

基　　金：广东省科技计划项目（2010B031600095）

摘　　要：目的探讨NLRP3（nucleotide—binding oligomerization domain-leucine—rich repeats contalning pynn domain3）／IL-1p、IL．18通路在哮喘小鼠肺支气管炎症的关系。方法C57BL／6小鼠随机分为正常对照组、NLRP3抑制剂格列本脲组和哮喘组，每组7只；格列本脲组和哮喘组采用卵蛋白致敏、诱导建立哮喘小鼠模型，格列本脲组每次雾化激发前30min腹腔注射格列本脲500mS／kg；造模后检测小鼠气道反应性、苏木精-伊红（HE）染色观察小鼠肺组织病理改变、计数支气管肺泡灌洗液（BALF）中白细胞总数及嗜酸细胞百分比、淋巴细胞百分比以了解各组小鼠的支气管炎症程度；采用Westernblot和RT．PCR检测小鼠肺组织NLRP3蛋白和mRNA的表达；酶联免疫吸附试验（ELISA）检测小鼠肺组织匀浆上清IL-1p、IL-18含量；并分别对IL-1p、IL-18与BALF中嗜酸性粒细胞百分比进行相关性分析。结果哮喘组、格列本脲组气道反应性均高于对照组（P〈0．05）。哮喘组BALF中白细胞总数、嗜酸粒细胞百分比及淋巴细胞百分比[（29．88±4．97）×10^4／ml、（21．67±4．83）％、（31．87±5．31）％]均明显高于格列本脲组[（17．30±1．19）×10^4／ml、（12．45±1．12）％、（17．16±0．97）％，P均〈0．05]及对照组[（9．74±2．88）×10^4／ml、（1．15±0．26）％、（10．66±1．83）％，P均〈0．05]，而格列本脲组比对照组高（P均〈O．05）。哮喘组、格列本脲组肺组织病理检查可见支气管周围较多炎症细胞浸润，但格列本脲组较哮喘组炎症浸润少，而对照组肺组织基本无炎症细胞浸润。哮喘组肺组织NLRP3蛋白及mRNA表达均高于格列本脲组、对照组（P均〈0．05）。哮喘组肺组织匀浆上清IL-1β（74．81±17．45）ps／mg及IL-18（426．94±76．05）ps／mg含量均高于对照组[（37．54±5．53）ps／mg，P〈0．05；（249．62±161．20）pg／mg，P〈0．05]。IL-1βObjective To verify the hypothesis that NLRP3-dependent IL-1β and IL-18 play a pathogenic role in mice with asthma. Methods A total of 21 C57BL/6 mice were randomly divided into 3 groups： the control group, NLRP3 inhibitor glyburine group and asthma group （n = 7）. A mouse model of allergic asthma was established by sensitization and challenge with ovalbumin for the mice in glyburine group and asthma group. Glyburine at 0.5 mg/g was given to the mice in glyburine group by intraperitoneal injection 30 minutes before inhaling ovalbumin. After induction of asthma, the airway responsiveness was measured and the pathological change in lung tissues was observed with H＆E staining. The counts of total leuko- cytes and the percentages of eosinophils and lymphocytes in bronchoalveolar lavage fluid （BALF） were cal- culated for evaluation of airway inflammation. The expressions of NLRP3 and its mRNA were identified by Western blot and RT-PCR, respectively. The concentrations of IL-1β and IL-18 in mouse lung homogenate supernatant were detected by ELISA. Then the correlations between IL-1β/IL-18 and the percentage of eo- sinophils in BALF was analyzed. Results The airway responsiveness of the mice in asthma and glyburine groups were significantly higher than those in control group （P〈0. 05）. The total cell numbers of leukocytes and the percentages of eosinophils and lymphocytes in BALF of the mice in asthma group were the highest a- mong three groups （ P〈0.05 ）, followed by the glyburine group （P〈0.05）. The airway inflammation could be detected in both group B and C, however, be alleviated by glyburine intervention. The expressions of NL- RP3 protein and mRNA in the mice with asthma were higher than those in the control group and the glyburine group （P〈0.05）. The concentrations of IL-10 and IL-18 were increased in the mice in asthma group as compared with those in the control group. The correlative analysis showed that there was a positive correlation between IL-1β/IL-18 and the percenta

关 键 词：NLRP3 IL-1Β IL-18 哮喘小鼠 

分 类 号：R5[医药卫生—内科学]