白花前胡丙素抗哮喘模型小鼠Th17/Treg失衡作用

Regulation of Praeruptorin C on the Imbalance of Th17/Treg in a Mouse Asthmatic Model

关键词:  白花前胡丙素  哮喘  调节性T细胞

Key words:  praeruptorin C  asthma  regulatory T cell

摘要:

目的 研究白花前胡丙素对哮喘模型小鼠体内Th17/Treg失衡的干预作用。方法 小鼠腹腔注射和雾化吸入卵清蛋白以建立气道炎性反应模型，并灌胃不同剂量的白花前胡丙素干预。模型结束后，动物肺功能仪分析气道高反应性；细胞计数器和Diff-Quick染色计数支气管灌洗液中白细胞总数及分类；酶联免疫吸附法检测血清或BALF中细胞因子和炎性介质的水平；苏木精-伊红(HE)染色法观察气道病理改变；流式细胞检测脾脏CD4+细胞数及分类；RT-PCR分析脾脏Foxp3 mRNA的表达。结果 与对照组比较，模型组呈现明显的气道炎性反应和气道高反应(P<0.05或P<0.01)；IgE、IL-17水平增加(P<0.01)，IL-10、TGF-β1水平降低(P<0.01)；CD4⁺CD25⁺Foxp3⁺细胞占总CD4⁺细胞的比例降低(P<0.01)；Foxp3 mRNA的表达减少(P<0.01)。与模型组比较，白花前胡丙素组气道炎症显著减轻，中、高剂量组气道反应在氯乙酰胆碱剂量>10 mg·kg^-1均显著受到抑制(P<0.05或P<0.01)；白花前胡丙素组IgE水平显著降低(P<0.01)，IL-10、TGF-β1水平显著增加(P<0.05或P<0.01)，中、高剂量组IL-17显著降低(P<0.01)；白花前胡丙素组显著增加CD4⁺CD25⁺Foxp3⁺细胞的比例(P<0.05或P<0.01)，并增加Foxp3 mRNA的表达(P<0.05或P<0.01)。结论 白花前胡丙素具有抗哮喘鼠Th17/Treg失衡的作用。

Abstract:

OBJECTIVE To study the effects of praeruptorin C on the imbalance of Th17/Treg in a mouse model of allergic asthma. METHODS BALB/c mice were sensitized and challenged by OVA to induce airway inflammation, and administered intragastrically with praeruptorin C at different doses. Airway responsiveness was measured by a lung function analysis systems. The number of total and differential leukocytes in BALF was counted using a hemocytometer or Diff-Quick-stained smears. The pathological inspection of lung tissue was analyzed by hematoxylin-eosin staining. Levels of cytokine and inflammatory mediators in BALFs or serum were measured by enzyme-linked immunosorbent assay. Numbers of CD4⁺ T cells in spleen was analyzed by flow cytometry. The expression of Foxp3 mRNA in spleen was detected by reverse transcription polymerase chain reaction. RESULTS Compared with control group, the model group exhibited obvious airway inflammation and hyperreactivity(P<0.05 or P<0.01), the levels of IgE and IL-17 were significantly increased(P<0.01), levels of IL-10 and TGF-β1 was significantly decreased(P<0.01), the percentages of CD4⁺CD25⁺Foxp3⁺ regulatory T cell among the CD4⁺ T cells were significantly decreased(P<0.01), and the expression of Foxp3 mRNA was significantly decreased(P<0.01). Compared with the model group, the group of praeruptorin C exhibited ease of airway inflammation, airway hyperreactivity in the middle and high dose of praeruptorin C groups were obviously suppressed as the dose of acetylcholine chloride were more than 10 mg·kg^-1, the groups of praeruptorin C reduced the levels of IgE significantly(P<0.01) and increased the levels of IL-10, TGF-β1(P<0.05 or P<0.01), the middle and high dose of praeruptorin C reduced the levels of IL-17 significantly(P<0.01), the groups of praeruptorin C increased the percentages of CD4⁺CD25⁺Foxp3⁺ regulatory T cell and the expression of Foxp3 mRNA(P<0.05 or P<0.01). CONCLUSION Praeruptorin C efficiently regulates the imbalance of Th17/Treg to balance a mouse model of allergic airway inflammation.