中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2010年
4期
303-308
,共6页
李艳丽%李鸿佳%亓慧娟%王荣%姬峰%郝俊青%毕文祥%董亮
李豔麗%李鴻佳%亓慧娟%王榮%姬峰%郝俊青%畢文祥%董亮
리염려%리홍가%기혜연%왕영%희봉%학준청%필문상%동량
气道上皮%TSLP%树突状细胞%支气管哮喘
氣道上皮%TSLP%樹突狀細胞%支氣管哮喘
기도상피%TSLP%수돌상세포%지기관효천
Airway epithelium%TSLP%Dendritic cell%Bronchial asthma
目的 研究支气管哮喘小鼠气道上皮中胸腺间质淋巴细胞生成素(TSLP)表达,探讨其对哮喘小鼠肺部炎症的影响.方法 BALB/c小鼠分为生理盐水对照组、哮喘模型组和TSLP中和抗体干预组.通过气道反应性和肺组织病理学评价哮喘模型;酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液(BALF)上清中IL-4、IL-5和IL13的含量;实时荧光定量PCR(qRT-PCR)测定肺组织中TSLP mRNA的表达;免疫组化及Western blot法测定肺组织中TSLP蛋白的表达;流式细胞术检测BALF中树突状细胞(OCs)表面CD40、CD80、CD86的表达水平.结果 小鼠气道反应性增高和肺组织病理学检查结果均符合哮喘的典型表现证实造模成功;哮喘组BALF中IL-4、IL-5和IL-13的水平显著高于正常组(P<0.05),且TSLP与其成正相关;与正常对照组相比,哮喘组气道上皮TSLPmRNA和蛋白高表达,两组间差异有统计学意义(P<0.05);哮喘组BALF中DCs表面CD40、CD80、CD86表达明显高于正常组(P<0.05).TSLP中和抗体干预后,BALF中DCs表面CD40、CD80、CD86表达明显减低,并进一步减少IL-4、IL-5和IL-13的表达.结论 哮喘气道上皮中TSLP表达增高,TSLP通过上调DCs表面CD40、CD80、CD86的表达,激活DCs诱导CD4~+T细胞向Th2分化发育,与加重哮喘的气道炎症有关;TSLP抗体干预可阻断DCs的活化,减少Th2细胞因子的分泌,这些因素可能与减轻哮喘炎症反应有关,为哮喘治疗途径提供新的思路.
目的 研究支氣管哮喘小鼠氣道上皮中胸腺間質淋巴細胞生成素(TSLP)錶達,探討其對哮喘小鼠肺部炎癥的影響.方法 BALB/c小鼠分為生理鹽水對照組、哮喘模型組和TSLP中和抗體榦預組.通過氣道反應性和肺組織病理學評價哮喘模型;酶聯免疫吸附試驗(ELISA)檢測支氣管肺泡灌洗液(BALF)上清中IL-4、IL-5和IL13的含量;實時熒光定量PCR(qRT-PCR)測定肺組織中TSLP mRNA的錶達;免疫組化及Western blot法測定肺組織中TSLP蛋白的錶達;流式細胞術檢測BALF中樹突狀細胞(OCs)錶麵CD40、CD80、CD86的錶達水平.結果 小鼠氣道反應性增高和肺組織病理學檢查結果均符閤哮喘的典型錶現證實造模成功;哮喘組BALF中IL-4、IL-5和IL-13的水平顯著高于正常組(P<0.05),且TSLP與其成正相關;與正常對照組相比,哮喘組氣道上皮TSLPmRNA和蛋白高錶達,兩組間差異有統計學意義(P<0.05);哮喘組BALF中DCs錶麵CD40、CD80、CD86錶達明顯高于正常組(P<0.05).TSLP中和抗體榦預後,BALF中DCs錶麵CD40、CD80、CD86錶達明顯減低,併進一步減少IL-4、IL-5和IL-13的錶達.結論 哮喘氣道上皮中TSLP錶達增高,TSLP通過上調DCs錶麵CD40、CD80、CD86的錶達,激活DCs誘導CD4~+T細胞嚮Th2分化髮育,與加重哮喘的氣道炎癥有關;TSLP抗體榦預可阻斷DCs的活化,減少Th2細胞因子的分泌,這些因素可能與減輕哮喘炎癥反應有關,為哮喘治療途徑提供新的思路.
목적 연구지기관효천소서기도상피중흉선간질림파세포생성소(TSLP)표체,탐토기대효천소서폐부염증적영향.방법 BALB/c소서분위생리염수대조조、효천모형조화TSLP중화항체간예조.통과기도반응성화폐조직병이학평개효천모형;매련면역흡부시험(ELISA)검측지기관폐포관세액(BALF)상청중IL-4、IL-5화IL13적함량;실시형광정량PCR(qRT-PCR)측정폐조직중TSLP mRNA적표체;면역조화급Western blot법측정폐조직중TSLP단백적표체;류식세포술검측BALF중수돌상세포(OCs)표면CD40、CD80、CD86적표체수평.결과 소서기도반응성증고화폐조직병이학검사결과균부합효천적전형표현증실조모성공;효천조BALF중IL-4、IL-5화IL-13적수평현저고우정상조(P<0.05),차TSLP여기성정상관;여정상대조조상비,효천조기도상피TSLPmRNA화단백고표체,량조간차이유통계학의의(P<0.05);효천조BALF중DCs표면CD40、CD80、CD86표체명현고우정상조(P<0.05).TSLP중화항체간예후,BALF중DCs표면CD40、CD80、CD86표체명현감저,병진일보감소IL-4、IL-5화IL-13적표체.결론 효천기도상피중TSLP표체증고,TSLP통과상조DCs표면CD40、CD80、CD86적표체,격활DCs유도CD4~+T세포향Th2분화발육,여가중효천적기도염증유관;TSLP항체간예가조단DCs적활화,감소Th2세포인자적분비,저사인소가능여감경효천염증반응유관,위효천치료도경제공신적사로.
Objective To study the expression of thymic stromal lymphopoietin(TSLP) and the activation of DCs in OVA-induced murine asthma model, and investigate the effects and underlying mecha-nisms of TSLP on lung inflammation. Methods Thirty BALB/c mice were randomly divided into control group, OVA group and TSLP neutralizing antibody treated group. The asthma model was evaluated by airway responsiveness and histological analysis of lung tissues ; The levels of TSLP mRNA in lungs were determined by quantitative real-time PCR; The expression of TSLP in lungs were determined by immunohistochemistry and Western blot; The expression of CD40, CD80, CD86 in BALF was detected by FACS. Results Both the histological analysis of lung tissues and the airway responsiveness were all consistent with the characteris-tic of murine asthma model. The expression of TSLP and TSLP mRNA in the OVA group was significantly in-creased compared with blank group. The expression of CD40, CD80, CD86 in BALF from OVA group was increased significantly compared with the control group. Furthermore, treating mice with TSLP neutralizing antibody reduced the expression of CD40, CD80, CD86 on dendritic cells, and IL-4, IL-5, IL-13 in the OVA group. Conclusion Our study indicate that TSLP was highly expressed in the bronchial epithelia of murine asthma model, via upregulation of CD40, CD80, CD86, induce DCs to active CD4~+ T cells and pro-duce type 2 responses, so that aggravating the lung inflammation of asthma. Blocking TSLP is capable of in-hibiting the production of Th2 cytokines, thus presents a promising strategy for the treatment of asthma.
