中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
16期
1219-1222
,共4页
何芳%沈启英%方磊%蒋旭琴%吴惠梅%刘荣玉
何芳%瀋啟英%方磊%蔣旭琴%吳惠梅%劉榮玉
하방%침계영%방뢰%장욱금%오혜매%류영옥
哮喘%高迁移率族蛋白质类%Toll样受体2%小鼠
哮喘%高遷移率族蛋白質類%Toll樣受體2%小鼠
효천%고천이솔족단백질류%Toll양수체2%소서
Asthma%High mobility group proteins%Toll-like receptor 2%Mice
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.
目的 探討Toll樣受體2(TLR2)介導的高遷移率族蛋白1(HMGB1)信號分子在小鼠支氣管哮喘(簡稱哮喘)中的作用機製.方法 非特定病原體(SPF)級雌性C57及TLR2-/-小鼠各14隻按隨機數字錶法分彆隨機分為2組共4組,即:C57對照組、C57哮喘組、TLR2基因缺失(TLR2-/-)對照組、TLR2-/-哮喘組,每組7隻.哮喘組採用卵清蛋白緻敏和霧化激髮模式建立哮喘小鼠模型,對照組以等量的生理鹽水代替.酶聯免疫吸附(ELISA)法測定支氣管肺泡灌洗液(BALF)上清中HMGB1含量.免疫組化及Western印跡法檢測肺組織HMGB1的錶達.結果 成功建立瞭小鼠哮喘模型.C57哮喘組BALF中HMGB1含量顯著高于C57對照組、TLR2-/-哮喘組、TLR2-/-對照組[(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].免疫組化檢測HMGB1在C57對照組及TLR2-/-對照組肺組織中少量錶達,而在C57哮喘組及TLR2-/-哮喘組中錶達明顯,併定位于氣道上皮中.Western印跡法檢測C57哮喘組肺組織中HMGB1相對錶達量顯著高于C57對照組、TLR2-/-哮喘組、TLR2-/-對照組(0.92±0.29比0.18±0.09、0.31±0.16、0.21±0.14,P=0.007、0.022、0.009).結論 TLR2介導的HMGB1信號分子促進瞭氣道的炎癥反應,可能參與瞭哮喘的髮病.
목적 탐토Toll양수체2(TLR2)개도적고천이솔족단백1(HMGB1)신호분자재소서지기관효천(간칭효천)중적작용궤제.방법 비특정병원체(SPF)급자성C57급TLR2-/-소서각14지안수궤수자표법분별수궤분위2조공4조,즉:C57대조조、C57효천조、TLR2기인결실(TLR2-/-)대조조、TLR2-/-효천조,매조7지.효천조채용란청단백치민화무화격발모식건립효천소서모형,대조조이등량적생리염수대체.매련면역흡부(ELISA)법측정지기관폐포관세액(BALF)상청중HMGB1함량.면역조화급Western인적법검측폐조직HMGB1적표체.결과 성공건립료소서효천모형.C57효천조BALF중HMGB1함량현저고우C57대조조、TLR2-/-효천조、TLR2-/-대조조[(59.0±13.9)비(42.3±1.6)、(47.5±2.3)、(42.4±1.4) ng/L,P=0.001、0.037、0.001].면역조화검측HMGB1재C57대조조급TLR2-/-대조조폐조직중소량표체,이재C57효천조급TLR2-/-효천조중표체명현,병정위우기도상피중.Western인적법검측C57효천조폐조직중HMGB1상대표체량현저고우C57대조조、TLR2-/-효천조、TLR2-/-대조조(0.92±0.29비0.18±0.09、0.31±0.16、0.21±0.14,P=0.007、0.022、0.009).결론 TLR2개도적HMGB1신호분자촉진료기도적염증반응,가능삼여료효천적발병.
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.
