HSP70/CD80 DNA疫苗通过调节趋化因子及受体对哮喘小鼠气道炎症的影响

目的 观察HSP70/CD80 DNA疫苗通过调节趋化因子及趋化因子受体对急性哮喘小鼠气道炎症和气道高反应性的治疗作用.方法 以卵清蛋白(OVA)致敏小鼠,建立小鼠急性哮喘模型,同时肌肉注射HSP70/CD80 DNA疫苗.观察小鼠气道反应性,外周血IgE,肺组织炎症和黏液分泌情况,CCL5和CCL17在气道表达情况.Real-time PCR检测肺组织CCR5和CCR4基因表达.结果 与对照小鼠相比,急性哮喘小鼠的气道反应性明显增高,血清IgE含量明显升高(P＜0.05),肺组织炎细胞浸润明显(P＜0.05),大量黏液形成.HSP70/CD80 DNA疫苗治疗小鼠后,能有效减轻气道高反应性,降低血清IgE含量,降低抑制气道炎细胞浸润,减少黏液分泌(P＜0.05);且CCL5在气道上皮呈阳性表达,CCL17呈阴性(P＜0.05);肺组织CCR5基因表达增加和CCR4表达减少(P＜0.05).结论 HSP70/CD80 DNA疫苗可通过增强CCL5在气道上皮中的表达,减少CCL17表达;上调CCR5抑制CCR4,使CCR5/CCR4升高,从而恢复Th 1/Th2平衡,起到治疗哮喘的作用.     

抗SH2-Bβ抗体可减轻哮喘小鼠气道炎症和气道高反应性

目的:研究SH2-Bβ在支气管哮喘发病中的作用.方法:SH2-Bβ被阻断后,观察哮喘小鼠肺泡灌洗液中细胞数、应用酶联免疫吸附实验肺泡灌洗液、血清及脾细胞培养液中IgE、IL-4含量变化.应用小鼠肺功能仪检测气道阻力的变化.结果:同哮喘组比,anti-SH2-Bβ组:1)气道阻力明显降低(P＜0.01);2)肺泡灌洗液中细胞总数及嗜酸细胞数明显降低(P＜0.01);3)血清IgE及脾细胞培养液中IgE、IL-4含量明显降低(P＜0.01).结论:SH2-Bβ参与哮喘小鼠气道炎症和气道高反应性的发病机制.阻断SH2-Bβ可能成为治疗过敏性哮喘的有效新方法.     

Th17淋巴细胞及相关细胞因子加重哮喘小鼠气道炎症

目的:研究Th17淋巴细胞及其相关的细胞因子在加重哮喘小鼠气道炎症中的作用机制.方法:20只小鼠随机均分为哮喘组和正常对照组.哮喘组用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型.对照组致敏与激发均以生理盐水代替.HE染色观察小鼠气道及肺组织病理变化;光学显微镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞分类及计数;酶联免疫吸附试验(ELISA)检测小鼠BALF上清中IL-4、IFN-γ及IL-17的含量,流式细胞技术(FCM)检测小鼠外周血Th1、Th2及Th17淋巴细胞占CD4+细胞百分率情况.将外周血各T淋巴细胞亚群与BALF的中性粒细胞数作相关性分析.结果:哮喘组小鼠BALF中细胞数及中性粒细胞、嗜酸性粒细胞、淋巴细胞百分率均显著高于对照组(P＜0.05),BALF上清中IL-4和IL-17的水平显著增高(P＜0.05),而IFN-γ的水平显著降低(P＜0.05),外周血Th2、Th17淋巴细胞占CD4+细胞百分比显著增高(P＜0.05),而Th1淋巴细胞占CD4+细胞百分比显著降低(P＜0.05).相关性分析显示Th1淋巴细胞与BALF的中性粒细胞数呈显著负相关(rThl=-0.446,P＜0.05),Th17淋巴细胞与BALF的中性粒细胞数呈显著正相关(rTh17=0.394,P＜0.05).结论:Th17淋巴细胞可促进中性粒细胞及炎性介质在哮喘小鼠气道内的聚集,加重气道炎症.     

Thl7淋巴细胞在哮喘小鼠气道炎症中的初步研究

目的:初步探索Th17细胞及其分泌的炎症介质在哮喘小鼠气道炎症中的作用机制.方法:20只小鼠随机均分为哮喘组和正常对照组.哮喘组用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型.正常对照组致敏与激发均以生理盐水代替.HE染色观察小鼠气道及肺组织病理变化;光学显微镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞分类及计数;酶联免疫吸附试验(ELISA)检测小鼠BALF上清中IL-4、IL-5、IL-13、IFN-γ及IL-17的含量,流式细胞技术(FCM)检测小鼠外周血Th1、Th2及Th17淋巴细胞占CD4+T淋巴细胞百分率情况.结果:哮喘组小鼠BALF中细胞总数和中性粒细胞、嗜酸性粒细胞、淋巴细胞百分率均显著高于对照组(P<0.05),BALF上清中IL-4、IL-5、IL-13及IL-17的水平显著增高(P<0.05),而IFN-γ差异无统计学意义(P>0.05),外周血Th2、Th17细胞明显增高(P<0.05),而Th1细胞无明显变化.结论:Th17细胞及其分泌的炎症介质可促进中性粒细胞及嗜酸性粒细胞在气道内聚集,加重哮喘气道炎症,可能与哮喘气道重塑密切相关.     

慢性尘螨暴露诱导小鼠气道Th1炎症相关的气道重塑

目的:观察慢性尘螨变应原暴露对小鼠气道变应性炎症及重构的影响。方法:采用α平滑肌肌动蛋白启动子驱动的Cre重组酶(α-SMA-Cre)与R26R双转基因报告小鼠,尘螨连续滴鼻60 d后测定气道阻力;肺泡灌洗并分类计数;分离肺组织并提取肺组织蛋白;制备病理组织切片;分离培养脾脏淋巴细胞。结果:慢性尘螨暴露组小鼠气道阻力明显升高(P0.01),肺泡灌洗液(BALF)中细胞总数和淋巴细胞比例显著高于正常对照组(P0.01),肺组织HE染色示:慢性尘螨暴露组小鼠气道上皮细胞炎症水肿,小血管周围淋巴细胞浸润,未见嗜酸粒细胞。X-gal染色显示:尘螨暴露组气道平滑肌细胞及上皮下肌纤维母细胞明显增生,相应Western blotting结果显示α平滑肌肌动蛋白(α-SMA)表达明显增多。尘螨暴露组肺匀浆上清γ干扰素(IFN-γ)较正常对照组升高(P0.01),白细胞介素-4(IL-4)水平两者间没有显著差异(P0.05)。流式细胞检测显示尘螨暴露组分泌IFN-γ的CD4+Th1细胞较正常对照组增多。结论:慢性尘螨气道暴露诱导了Th1炎症,与气道重构和气道高反应性密切相关。     

哮喘小鼠肺组织中转录因子RORγt的表达与气道炎症的关系

目的:探讨Th17细胞转录因子RORγt在支气管哮喘小鼠肺组织中的表达及其与哮喘气道炎症的关系.方法:采用卵清蛋白(OVA)致敏方法建立支气管哮喘小鼠模型;BALB/c小鼠30只随机分为对照组、哮喘组、地塞米松治疗组各10只.采用酶联免疫吸附试验(ELISA)方法检测小鼠肺泡灌洗液(BALF)、血清中白细胞介素17(IL-17)水平;HE染色评价各组小鼠气道炎症情况;逆转录聚合酶链反应(RT-PCR)方法检测肺组织IL-17、RORγt mRNA表达水平;免疫印记(Western blot)方法检测肺组织RORγt蛋白表达水平.结果:哮喘组小鼠肺组织RORγt mRNA、蛋白水平及IL-17水平均明显高于对照组和地塞米松治疗组(P＜0.05),RORγt蛋白表达量与嗜酸性粒细胞数、淋巴细胞数、中性粒细胞数、BALF、外周血中IL-17含量、肺组织IL-17 mRNA表达量均呈正相关关系(r＝0.789、0.795、0.902、0.669、0.806、0.883,P值均＜0.01).结论:RORγt在支气管哮喘小鼠肺组织呈高表达,其表达水平与气道炎症密切相关,参与了哮喘气道炎症的发生过程.     

哮喘小鼠气道上皮TSLP表达及激活DCs加重气道炎症的研究

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.     

IL-27对哮喘小鼠气道炎症的影响

目的研究IL-27对卵白蛋白(OVA)激发哮喘小鼠气道炎症的影响。方法 24只雌性BALB/c小鼠随机分为生理盐水组、哮喘组及IL-27组,每组8只。应用OVA建立哮喘模型,IL-27组小鼠应用1μgIL-27(溶于50μlPBS中)滴鼻给药,观察3组小鼠肺组织病理改变,计数支气管肺泡灌洗液(BALF)中嗜酸性粒细胞;ELISA法测定小鼠BALF中IL-4和IFN-γ浓度,RT-PCR测定肺组织T-bet mRNA的表达量。结果 IL-27组小鼠肺组织炎症反应明显轻于哮喘组小鼠;IL-27组小鼠BALF中嗜酸性粒细胞计数为(2.21±0.33)×107/L明显低于哮喘组的(12.82±2.17)×107/L(P0.01);IL-27组小鼠BALF中IL-4浓度为(20.4±3.2)μg/L,明显低于哮喘组的(61.3±13.1)μg/L(P0.05);IL-27组小鼠BALF中IFN-γ浓度为(50.3±6.3)μg/L,明显高于哮喘组的(11.1±3.3)μg/L(P0.05);IL-27组小鼠肺组织T-bet mRNA表达量(吸光度积分比值)为(0.268±0.048),明显高于哮喘组的(0.130±0.012)(P0.05)。结论 IL-27可能通过增强T-bet mRNA的表达增强Th1反应,减少BALF中嗜酸性粒细胞数量,进而减轻了哮喘小鼠肺组织炎症反应。     

HDM通过肺泡巨噬细胞TLR4诱导哮喘小鼠气道炎症的机制研究

目的:研究HDM通过肺泡巨噬细胞(AM)Toll样受体4(TLR4)的高表达及诱导AM的活化,探讨其对哮喘小鼠气道炎症的影响.方法:BALB/c小鼠随机分为哮喘模型组(OVA)A,HDM处理组(HDM+OVA)B,对照组(生理盐水)C.用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型;HE染色观察小鼠气道及肺组织病理变化;光学显微镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞分类及计数;酶联免疫吸附试验(ELISA)检测小鼠BALF上清中IL-4、IL-5、IL-13和IFN-γ的含量,实时定量PCR法测定AM的TLR4的表达,流式细胞技术(FCM)检测AM的CD80、CD86的表达.结果:与A组相比,B组BALF上清中IL-4、IL-5和IL-13的水平显著增高(P<0.05),而IFN-γ差异无统计学意义(P>0.05),与A组相比,AM的TLR4mRNA表达明显增高(P<0.05),CD80的表达差异无统计学意义,CD86的表达水平显著增高,差异有统计学意义(P<0.05).结论:HDM通过AM的TLR4的高表达诱导AM的活化,加重哮喘的气道炎症.     

Th17淋巴细胞在哮喘小鼠气道炎症中的初步研究

目的:初步探索Th17细胞及其分泌的炎症介质在哮喘小鼠气道炎症中的作用机制。方法:20只小鼠随机均分为哮喘组和正常对照组。哮喘组用卵白蛋白(OVA)致敏与激发建立小鼠哮喘模型。正常对照组致敏与激发均以生理盐水代替。HE染色观察小鼠气道及肺组织病理变化;光学显微镜下观察小鼠支气管肺泡灌洗液(BALF)中细胞分类及计数;酶联免疫吸附试验(ELISA)检测小鼠BALF上清中IL-4、IL-5、IL-13、IFN-γ及IL-17的含量,流式细胞技术(FCM)检测小鼠外周血Th1、Th2及Th17淋巴细胞占CD4+T淋巴细胞百分率情况。结果:哮喘组小鼠BALF中细胞总数和中性粒细胞、嗜酸性粒细胞、淋巴细胞百分率均显著高于对照组(P<0.05),BALF上清中IL-4、IL-5、IL-13及IL-17的水平显著增高(P<0.05),而IFN-γ差异无统计学意义(P>0.05),外周血Th2、Th17细胞明显增高(P<0.05),而Th1细胞无明显变化。结论:Th17细胞及其分泌的炎症介质可促进中性粒细胞及嗜酸性粒细胞在气道内聚集,加重哮喘气道炎症,可能与哮喘气道重塑密切相关。     

OVA-Fc融合基因疫苗对哮喘小鼠的疗效观察

目的探讨OVA-Fc融合基因疫苗治疗哮喘小鼠气道炎症和气道高反应性的效果。方法分别将制备的OVA-Fc-pcDNA3．1质粒、OVA-pcDNA3．1质粒、pcDNA3．1质粒,皮下免疫Balb／c哮喘小鼠模型,40d后,观察肺组织的病理变化,并检测肺泡灌洗液中细胞总数及嗜酸粒细胞计数,细胞因子的含量以及血清中OVA特异的IgE抗体的水平。结果OVA-Fc-pcDNA3．1基因修饰的DNA疫苗免疫小鼠后,能有效抑制哮喘小鼠肺组织的炎细胞浸润,肺泡灌洗液中细胞总数明显下降（P〈0．05）,嗜酸性粒细胞的总数明显下降（P〈0．05）,肺泡灌洗液中IL-10、INF-Y的分泌增加（P〈0．05）,血清中OVA特异的IgE抗体能有效的得以控制（P〈0．05）。结论OVA-Fc-pcDNA3．1基因修饰的DNA疫苗可较OVA变应原基因更强的特异性地抑制哮喘小鼠的气道炎症,有望对哮喘的治疗具有重要的价值。     

T-bet基因转染对哮喘小鼠气道炎症的影响

目的： 观察气道内T-bet基因转染对哮喘小鼠气道炎症的影响。方法: C57BL/6小鼠40只,随机分为4组,每组10只,分别为正常对照组（A组）、哮喘模型组（B组）、空质粒干预组(C组) 和T-bet质粒干预组(D组)。卵白蛋白(OVA) 抗原溶液腹腔注射致敏,滴鼻造模。正常对照组用生理盐水代替OVA,空质粒干预组和T-bet质粒干预组OVA激发48 h前,分别经鼻滴入50 μg空质粒和重组T-bet质粒。观察各组实验小鼠的肺组织炎症以及BALF中各类炎症细胞以及IL-4、IFN-γ水平的变化。 结果: Western blotting检测发现,小鼠气道转染pcDNA3-T-bet质粒48 h后肺组织T-bet蛋白表达显著增加。pcDNA3-T-bet质粒转染能较好抑制给药后48 h OVA激发的哮喘小鼠气道炎症（包括炎症细胞浸润,上皮细胞损伤、黏液分泌、血管壁水肿及管腔缩窄）;下调小鼠BALF中Th2因子IL-4并上调Th1因子IFN-γ水平。 结论: 气道内转染T-bet质粒能有效改善哮喘小鼠的气道炎症。     

雾化吸入灭活草分支杆菌减轻哮喘小鼠气道炎症及对Toll样受体2表达的影响

目的研究雾化吸入灭活草分支杆菌对支气管哮喘小鼠气道炎症及肺组织细胞因子分泌的影响,探讨Toll样受体2(TLR2)表达在雾化吸入灭活草分支杆菌防治支气管哮喘中的作用。方法将24只雄性Balb/c小鼠按随机数字表法分为3组,每组8只:正常对照组(A)、哮喘模型组(B)、干预组(C)。卵清蛋白(OVA)致敏制小鼠支气管哮喘模型。C组在每次卵蛋白激发前给予雾化吸入草分枝杆菌治疗,每天1次。各组动物处死后提取肺组织和支气管肺泡灌洗液(BALF)。进行病理HE染色、AB-PAS染色观察支气管肺炎症和气道粘液分泌情况,并行半定量分析。BALF中炎症细胞计数,检测BALF中IL-4、IL-10、IFN-γ水平。实时定量PCR检测肺组织TLR2 mRNA表达水平。结果干预组BALF中IL-4分泌减少,IL-10、IFN-γ增加(P<0.05),BALF中嗜酸性粒细胞比例低于模型组,气道炎症病变较模型组减轻,肺组织TLR2 mRNA表达水平较模型组显著升高(P<0.05)。结论吸入草分枝杆菌能减轻支气管哮喘小鼠气道炎症,其效应与调节肺内细胞因子分泌有关。草分枝杆菌可能通过上调TLR2基因的表达调节支气管哮喘的免疫失衡。     

Prolonged antigen exposure ameliorates airway inflammation but not remodeling in a mouse model of bronchial asthma.

BACKGROUND: In naive rodents, repeated exposure to aerosolized antigen induces suppression of the Th2 response to the antigen. We hypothesized that more prolonged exposure of established asthma model to antigen aerosols may downregulate asthmatic phenotype. METHODS: After establishing an ovalbumin (OVA)-induced asthma model, mice were further exposed to OVA (prolonged exposure group) or phosphate-buffered saline (positive controls) 3 days per week for 6 weeks. During week 7, the mice of both groups were finally challenged with OVA. RESULTS: Prolonged OVA exposure resulted in marked suppression of serum OVA-specific immunoglobulin E (IgE) antibody levels, eosinophilia of the airway, and airway hyperresponsiveness (AHR). However, airway remodeling characterized by goblet cell hyperplasia and airway fibrosis was observed to the same degree in both groups. These effects were accompanied by diminished production of Th2 cytokines such as interleukin-4 (IL-4), IL-5 and IL-13 in bronchoalveolar lavage fluid (BALF) and cultured supernatant of splenocytes. Furthermore, prolonged exposure markedly increased IL-12 levels in BALF. CONCLUSIONS: Prolonged antigen exposure has inhibitory effects on eosinophilic inflammation, AHR and IgE response to antigen, but not on airway remodeling, presumably via inhibition of Th2 cytokines and increased IL-12 production in the lungs.     

Critical role of CD23 in allergen-induced bronchoconstriction in a murine model of allergic asthma.

CD23-deficient and anti-CD23 monoclonal antibody-treated mice were used to investigate the role of the low-affinity receptor for IgE (CD23) in allergic airway inflammation and airway hyperresponsiveness (AHR). While there were no significant differences in ovalbumin (OVA)-specific IgE titers and tissue eosinophilia, evaluation of lung function demonstrated that CD23-/- mice showed an increased AHR to methacholine (MCh) when compared to wild-type mice but were completely resistant to the OVA challenge. Anti-CD23 Fab fragment treatment of wild-type mice did not affect the MCh-induced AHR but significantly reduced the OVA-induced airway constriction. These results imply a novel role for CD23 in lung inflammation and suggest that anti-CD23 Fab fragment treatment may be of therapeutic use in allergic asthma.     

Alveolar macrophage-derived vascular endothelial growth factor contributes to allergic airway inflammation in a mouse asthma model

Vascular endothelial growth factor (VEGF) is a potent proangiogenic factor that correlates with vascular permeability and remodelling in asthma. Recently, alveolar macrophages (AM) were shown to be an important source of VEGF during lung injury. Our previous studies demonstrated that AM are an important subset of macrophages in the initiation of asthmatic symptoms. Here, we further investigated whether AM-derived VEGF was required for allergic airway inflammation in asthma. In this study, we reported that the expression of VEGF in AM was significantly increased after allergen challenge. Depleting AM or neutralizing VEGF in alveolus prevented ovalbumin (OVA)-induced asthma-related inflammation by inhibiting the infiltration of inflammatory cells in the lung, reduced the level of the cytokines, IL-4, IL-5, and IL-13, in the bronchoalveolar lavage fluid (BALF) and decreased airway hyperresponsiveness (AHR). Moreover, the inhibition of miR-20b increased the protein level of VEGF in normal AM; conversely, increasing miR-20b in asthmatic AM resulted in decreased VEGF protein levels. These findings suggest that AM-derived VEGF is necessary for allergic airway inflammation in asthmatic mice and miR-20b negatively regulates this expression.     

Time sequence of airway remodeling in a mouse model of chronic asthma: the relation with airway hyperresponsiveness

During the course of establishing an animal model of chronic asthma, we tried to elucidate the time sequence of airway hyperresponsiveness (AHR), airway inflammation, airway remodeling, and associated cytokines. Seven-week-old female BALB/c mice were studied as a chronic asthma model using ovalbumin (OVA). After sensitization, mice were exposed twice weekly to aerosolized OVA, and were divided into three groups depending on the duration of 4 weeks, 8 weeks, and 12 weeks. At each time point, airway responsiveness, inflammatory cells, cytokines in bronchoalveolar lavage fluids (BALF), serum OVA-specific IgE, IgG1, IgG2a, and histological examination were carried out. AHR to methacholine, increased levels of OVA-specific IgG1 and IgG2a, and goblet cell hyperplasia were continuously sustained at each time point of weeks. In contrast, we observed a time-dependent decrease in serum OVA-specific IgE, BALF eosinophils, BALF cytokines such as IL-13, transforming growth factor-beta1, and a time-dependent increase in BALF promatrix metalloproteinase-9 and peribronchial fibrosis. In this OVA-induced chronic asthma model, we observed airway remodelings as well as various cytokines and inflammatory cells being involved in different time-dependent manners. However, increased airway fibrosis did not directly correlate with a further increase in airway hyperresponsiveness.     

Intranasal challenge with increasing ovalbumin doses differently affects airway hyperresponsiveness and inflammatory cell accumulation in mouse model of asthma

Objective

To investigate whether challenge with increasing allergen doses could differently affect allergen-induced airway hyperresponsiveness (AHR) and inflammatory cell accumulation in mouse model of asthma, providing an experimental model to investigate their relationship.

Material and methods

AHR and accumulation of inflammatory cells in bronchoalveolar lavage fluid (BALF) and into the lungs were compared in ovalbumin-sensitized mice that were challenged intranasally with 2.5, 10, 25 or 100 μg of ovalbumin/mouse.

Results

Both AHR and inflammatory cell accumulation were proportional to the ovalbumin dose used for challenge. However, in group challenged with 10 μg of ovalbumin airway inflammation was present, although allergen-induced AHR was not detected. Additional analysis indicated that neither mucous hyperproduction nor eosinophil degranulation could be correlated to presence of AHR in this model, whereas concentration of interleukin (IL)-13 in BALF was increased only in those groups in which AHR was present.

Conclusions

Altogether, intranasal challenge of mice with increasing allergen doses could serve as a suitable experimental system for investigation of mechanisms by which airway inflammation leads to allergen-induced AHR. Our initial findings are in line with previous reports that dissociate AHR from amount of eosinophil accumulation and imply the role of IL-13 in this process.     

T-bet质粒基因转染对哮喘小鼠支气管肺泡灌洗液白细胞介素13和内皮素1的影响

目的 观察气道内T-bet质粒基因转染对哮喘小鼠气道肺泡灌洗液白细胞介素13(IL13)、内皮素1(ET-1)的影响.方法 C57BL/6J小鼠32只,完全随机分为4组(n=8),即哮喘模型组(A组)、正常对照组(B组)、空质粒干预组(C组)和模型T-bet质粒干预组(D组).用卵白蛋白(OVA)抗原溶液小鼠腹腔注射致敏,滴鼻造模.B组用生理盐水代替卵白蛋白,C组和D组卵白蛋白激发48 h前,分别经鼻滴入50μg空质粒或重组T-bet质粒.免疫印迹检测和酶联免疫吸附法(ELISA)检测各组小鼠的支气管肺泡灌洗液中IL-13、ET-1的水平.结果 哮喘模型组小鼠支气管肺泡灌洗液(BALF)中Th2因子IL-13和ET-1水平比正常对照组升高[(2.32±0.40)ng/L比(0.38±0.16)ng/L,(40.07±6.52)ng/L比(14.16±0.51)ng/L,均P＜0.05];模型T-bet质粒干预组BALF中Th2因子IL-13和ET-1水平[(1.16±0.19)和(23.08±2.59)ng/L]比哮喘模型组降低(均P＜0.05).结论 气道内转染T-bet质粒能有效下调小鼠BALF中Th2因子IL-13及ET-1水平.