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目的:探讨CD39和CD73阳性的CD4+CD25+Foxp3+T淋巴细胞(Treg细胞)对支气管哮喘(简称哮喘)小鼠气道炎症的影响。方法:将16只BALB/c成年雌性小鼠随机分为哮喘组(n=8)和对照组(n=8)。用卵清蛋白进行哮喘造模后,取小鼠血清检测总Ig E,采用高效液相色谱法行支气管肺泡灌洗液上清液三磷酸腺苷(adenosine-triphosphate,ATP)检测,取左肺组织行苏木精-伊红(hematoxylin eosin,HE)染色,观察小鼠肺组织的炎症改变;右肺上叶行CD39 m RNA、CD73 m RNA、Foxp3 m RNA检测;余右肺制成单细胞悬液,采用流式细胞术检测CD39+Treg和CD73+Treg细胞数量,计算其占Treg细胞百分比。结果 :哮喘组小鼠的血清总Ig E较对照组显著上升(P<0.01),支气管肺泡灌洗液中ATP较对照组升高(P<0.05),病理组织切片检查发现肺部炎症改变明显。哮喘组小鼠肺组织中CD39m RNA、CD73 m RNA、Foxp3 m RNA表达量分别为(0.54±0.11)×10-3、(1.11±0.36)×10-3、(0.08±0.03)×10-3,对照组则分别为(0.88±0.25)×10-3、(1.88±0.37)×10-3、(0.11±0.02)×10-3,2组间差异有统计学意义(P均<0.05)。流式细胞术检测发现,哮喘组小鼠肺组织的CD39+Treg、CD73+Treg细胞百分比较对照组略有下降趋势,但差异尚无统计学意义。结论:肺局部CD39+Treg和CD73+Treg细胞数量减少和(或)功能障碍导致的细胞外ATP清除率降低,可能是哮喘气道炎症发生的重要机制之一。
Objective: To investigate the effect of CD39 and CD73-positive CD4 + CD25 + Foxp3 + T lymphocytes (Treg cells) on airway inflammation in bronchial asthma (asthma) mice. Methods: Sixteen BALB / c adult female mice were randomly divided into asthma group (n = 8) and control group (n = 8). After ovalbumin was used for asthma modeling, total serum IgE was detected in mouse serum, and adenosine-triphosphate (ATP) was detected by high performance liquid chromatography in bronchoalveolar lavage fluid. The hematoxylin eosin (HE) staining was used to observe the inflammatory changes in the lung tissue of mice. The right upper lung was examined by CD39 m RNA, CD73 m RNA and Foxp3 m RNA. The number of CD39 + Treg and CD73 + Treg cells was detected by flow cytometry and the percentage of Treg cells was calculated. Results: The total serum IgE of asthmatic mice increased significantly (P <0.01), the level of ATP in bronchoalveolar lavage fluid increased (P <0.05), and the pathological changes of pulmonary inflammation changed obviously . The expression of CD39mRNA, CD73mRNA and Foxp3mRNA in asthmatic mice were (0.54 ± 0.11) × 10-3, (1.11 ± 0.36) × 10-3, (0.08 ± 0.03) × 10-3 (0.88 ± 0.25) × 10-3, (1.88 ± 0.37) × 10-3, (0.11 ± 0.02) × 10-3 in the control group, there was significant difference between the two groups (all P <0.05) . The results of flow cytometry showed that the percentages of CD39 + Treg and CD73 + Treg cells in lung tissue of asthmatic mice decreased slightly compared with the control group, but the difference was not statistically significant. Conclusion: The decrease of extracellular ATP clearance caused by the decrease of the number of CD39 + Treg and CD73 + Treg cells and / or dysfunction of lung may be one of the important mechanisms of airway inflammation in asthma.