Toll样受体2介导的高迁移率族蛋白1信号分子在小鼠支气管哮喘中的作用机制  

Mechanism of signal molecule high mobility group box protein 1 mediated by Toll-like receptor 2 in murine asthma

作　　者：何芳[1] 沈启英[1] 方磊[1] 蒋旭琴[1] 吴惠梅[1] 刘荣玉[1] 

机构地区：[1]安徽医科大学第一附属医院干部呼吸内科安徽省老年病研究所安徽省老年病分子医学重点实验室,合肥230022

出　　处：《中华医学杂志》2014年第16期1219-1222,共4页National Medical Journal of China

基　　金：国家自然科学基金（81270082、81170030）;高等学校博士学科点专项科研基金（20113420110006）;安徽省重点实验室计划项目（1206c0805028）;安徽省科技攻关项目（12010402135）

摘　　要：目的 探讨Toll样受体2（TLR2）介导的高迁移率族蛋白1（HMGB1）信号分子在小鼠支气管哮喘（简称哮喘）中的作用机制.方法 非特定病原体（SPF）级雌性C57及TLR2-/-小鼠各14只按随机数字表法分别随机分为2组共4组,即：C57对照组、C57哮喘组、TLR2基因缺失（TLR2-/-）对照组、TLR2-/-哮喘组,每组7只.哮喘组采用卵清蛋白致敏和雾化激发模式建立哮喘小鼠模型,对照组以等量的生理盐水代替.酶联免疫吸附（ELISA）法测定支气管肺泡灌洗液（BALF）上清中HMGB1含量.免疫组化及Western印迹法检测肺组织HMGB1的表达.结果 成功建立了小鼠哮喘模型.C57哮喘组BALF中HMGB1含量显著高于C57对照组、TLR2-/-哮喘组、TLR2-/-对照组[（59.0±13.9）比（42.3±1.6）、（47.5±2.3）、（42.4±1.4） ng/L,P=0.001、0.037、0.001].免疫组化检测HMGB1在C57对照组及TLR2-/-对照组肺组织中少量表达,而在C57哮喘组及TLR2-/-哮喘组中表达明显,并定位于气道上皮中.Western印迹法检测C57哮喘组肺组织中HMGB1相对表达量显著高于C57对照组、TLR2-/-哮喘组、TLR2-/-对照组（0.92±0.29比0.18±0.09、0.31±0.16、0.21±0.14,P=0.007、0.022、0.009）.结论 TLR2介导的HMGB1信号分子促进了气道的炎症反应,可能参与了哮喘的发病.Objective To explore the role and mechanism of signal molecule high mobility group box protein 1 （HMGB1） mediated by Toll-like receptor 2 （TLR2） in a murine asthma model.Methods Fourteen specific pathogen free （SPF） female C57 and TLR2 / mice each were randomly divided into 4 groups of C57 control,C57 asthma,TLR2-/-control and TLR2 / asthma （n =7 each）.The animals were sensitized and challenged with ovalbumin （OVA） for asthmatic modeling.The same amount of normal saline was used in the control group.The supernatant of bronchoalveolar lavage fluid （BALF） was collected for detecting the level of HMGB1 by enzyme-linked immunosorbent assay （ELISA）.And the expression of HMGB1 in lung tissue was detected by Western blot and immunohistochemistry.Results Asthmatic murine model was successfully established.The level of HMGB1 in the BALF of Cs7 asthma group was significantly higher than that in C57 control,TLR2-/ asthma and TLR2-/-control groups （（59.0 ± 13.9） vs （42.3 ± 1.6）,（47.5 ± 2.3）,（42.4 ± 1.4） ng/L; P =0.001,0.001,0.037） The results of immunohistochemistry showed that the marker of HMGB1 in lung tissue was less than those in the C57 control and TLR2-/-control groups.However,the C57 asthma and TLR2-/-asthma groups were obviously more and they were located in airway epithelium.Western blot showed that the expression of HMGB1 was significantly higher in C57 asthma group than that in the C57 control,TLR2-/-asthma and TLR2-/-control groups （0.92±0.29 vs 0.18±0.09,0.31 ±0.16,0.21 ±0.14; P =0.007,0.022,0.009）.Conclusions HMGB1 promotes the airway inflammation mediated by TLR2.And it may participate in the pathogenesis of asthma.

关 键 词：哮喘 高迁移率族蛋白质类 TOLL样受体2 小鼠 

分 类 号：R562.25[医药卫生—呼吸系统]