慢性阻塞性肺疾病大鼠体内调节性T细胞、Th17细胞的变化

Changes of regulatory T cells,Th17 cells in rats with chronic obstructive pulmonary disease

目的观察慢性阻塞性肺疾病(COPD)模型大鼠调节性T细胞(Treg)、Th17细胞的变化。方法将30只大鼠随机均分为正常组、模型组。模型组采用烟熏加脂多糖(LPS)气管滴入方法建立COPD肺气虚证大鼠模型,模型复制成功第28天,采用动物肺功能仪检测大鼠肺功能,酶联免疫吸附法检测大鼠血清白细胞介素(IL)-1β、IL-10、IL-17、IL-35水平,免疫组化法检测肺组织FoxP3、IL-17表达,流式细胞术检测外周血Treg表达。结果模型组大鼠肺泡腔及肺间质内大量炎性细胞浸润,肺组织结构破坏。与正常组比较,模型组大鼠肺功能参数0.3 s用力呼气容积(FEV0.3)、用力肺活量(FVC)、FEV0.3/FVC值降低(P<0.01,P<0.05);模型组大鼠血清IL-1β、IL-17、肺组织IL-17表达升高,血清IL-10、IL-35、肺组织FoxP3、外周血CD4+CD25+Treg表达降低(P<0.01,P<0.05);相关性分析显示,肺功能参数FEV0.3、FEV0.3/FVC、最大呼气流量(PEF)分别与IL-1β、IL-17呈负相关,与IL-35、FoxP3及CD4+CD25+Treg呈正相关(P<0.05)。结论 COPD存在高炎症反应,CD4+CD25+Treg、Th17表达失衡,调节功能降低,导致炎症反应升高。

Objective To observe the change of regulatory T cells （Treg） and Thl7 cells in rat model of chronic obstructive pulmonary disease （COPD）. Methods 30 rats were randomly divided into normal group, and model group. In addition to the normal group, the remaining rats were smoked plus lipopolysaccharide （LPS） tracheal instillation method to establish COPD lung deficiency rat model. 28 days after the model copied, pulmonary function was tested using animals seized spirometer, intedeukin （IL）-1β, IL-10, IL-17, IL-35 in serum were detected by enzyme-linked immunosorbent assay, FoxP3, 1L-17 was detected in lung tissue by immunohistochemistry staining. Treg of peripheral was detected by flow cytometry. Results A large number of inflammatory cell infiltration and pulmonary alveolar interstitial intrinsic in rat model group. Compared with normal group, lung function parameters such as FEV0.3, FVC, FEV0.3/FVC were decreased in model group （P 〈0. 01 ,P 〈0. 05） ; IL-1β, IL-17 were increased, expression of IL-17 in serum, IL-10, IL-35, FoxP3, CD4^ ＋ CD25^ ＋ Treg were reduced （P 〈0.01 ,P 〈 0. 05）. Correlation analysis showed that lung function parameters FEV0.3, FEV0.3/FVC, PEF, and IL-1β, IL-17 were negative correlattion respectively, IL-35, FoxP3 and CD4^ ＋ CD25^＋ Treg were positive correlation （ P 〈 0. 05 ）. Conclusion There is high inflammatory response in COPD. The high inflammatory response is caused by the imbalanced expression of CD4 ^＋ CD25^ ＋ Treg and Th17, which leads to increase in inflammatory response.