摘要
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(P<0.01),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(P<0.01),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(P<0.01),白细胞介素-4(IL-4)水平两者间没有显著差异(P>0.05)。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。
AIM： To investigate the inflammatory characteristics in the airway of mice with chronic exposure to dust mite.METHODS： The α-SMA-Cre/R26R transgenic reporter mice were intranasally exposed to dust mite extract for 60 d（DME group）,and then subjected to the measurement of lung resistance.The performance of bronchoalveolar lavage,pathological changes of the lung tissues and splenocytes isolation 24 h after the last challenge were observed.The protein extracts from the lungs were subjected to the detection of α-smooth muscle actin（α-SMA） by Western blotting.The supernatants of the lung homogenate were collected for testing the levels of interleukin-4（IL-4） and interferon-γ（IFN-γ） with enzyme-linked immunosorbent assay.CD4＋ T-cell subsets of the splenocytes were analyzed by flow cytometry.RESULTS： The mice chronically exposed to dust mite extract demonstrated severe airway hyperresponsiveness.The pulmonary pathological sections with HE staining manifested strong evidence of airway remodeling in DME group,corresponding to an enhanced X-gal staining that is related to α-SMA activation in the subepithelial basement membrane of bronchia.Total cell and lymphocyte counts were increased in the lungs of DME group compared to control group.No difference was found in eosinophil count of mice between DME and control groups.There was an elevated level of IFN-γ in the lungs of DME challenged mice coordinated with an increased proportion of IFN-γ-producing CD4＋ T cells in the splenocytes.CONCLUSION： Chronic exposure to dust mite in the mice induces Th1-dominant inflammation with an airway hyperresponsiveness and the development of airway remodeling.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第8期1559-1564,共6页
Chinese Journal of Pathophysiology
基金
国家科技部863重大资助项目(No.2007AA022002)
广东省自然科学基金重点资助项目(No.07117506)
