[关键词]
[摘要]
【目的】观察金丝桃苷对过敏性哮喘小鼠辅助性T (Th) 9、Th17、调节性T (Treg) 细胞分化及气道炎症的影响。 【方法】将60只雄性BALB/c小鼠随机分为5组,即正常组,模型组,金丝桃苷低、高剂量组,地塞米松组,每组12只。除正常组,其他各组小鼠采用卵清蛋白 (OVA) 诱导法构建过敏性哮喘模型。给药结束后,观测吸气阻力 (Ri) 、呼气阻力 (Re) 和肺动态顺应性 (Cldyn) ,评估肺功能,采用苏木素-伊红 (HE) 以及高碘酸希夫 (PAS) 染色法评估肺部组织病理学变化,流式细胞术分析脾脏Th9细胞、Th17细胞和Treg细胞比例,采用酶联免疫吸附分析 (ELISA) 检测血清和支气管肺泡灌洗液 (BALF) 中白细胞介素 (IL) -9、IL-10、IL-17A和转化生长因子 (TGF) -β1水平,采用实时定量聚合酶链反应 (PCR) 法检测肺组织中IL-9、IL-10、IL-17A和TGF-β1 mRNA表达水平,通过免疫组织化学法和Western Blot法检测肺组织中PU.1、维甲酸相关孤核受体γt (RORγt) 和叉头状转录因子3 (Foxp3) 蛋白表达水平。 【结果】与模型组比较,金丝桃苷高剂量组小鼠哮喘发作症状减轻,Ri和Re显著降低、Cldyn显著升高,BALF中白细胞总数、EOS数量显著降低,Th9和Th17细胞比例降低、Treg细胞比例升高,IL-9和IL-17A在肺组织中的mRNA水平和在血清、BALF中的含量显著降低,IL-10和TGF-β1在肺组织中的mRNA水平和在血清、BALF中的含量显著升高,肺组织中PU.1和RORγt蛋白表达水平显著降低,Foxp3蛋白表达水平显著升高,差异均有统计学意义 (P<0.05) 。 【结论】金丝桃苷可抑制过敏性哮喘小鼠Th9、Th17细胞分化、促进Treg细胞分化,减轻气道炎症。
[Key word]
[Abstract]
Objective To observe the effect of hyperoside on the differentiation of helper T (Th)9, Th17,regulatory T(Treg) cells and airway inflammation in allergic asthmatic mice. Methods Sixty male BALB/c mice were randomly divided into five groups,namely normal group,model group,hyperoside low- and high- dose groups,and dexamethasone group,with 12 mice in each group. Except for the normal group,the allergic asthma model was constructed using the ovalbumin (OVA) induction method in all groups of mice. At the end of drug administration, inspiratory resistance (Ri), exhalation resistance(Re) and pulmonary dynamic compliance (Cldyn) were observed to assess pulmonary function,hematoxylin-eosin (HE) and periodic acid Schiff (PAS) staining were used to assess histopathological changes in the lung tissues,and the ratios of splenic Th9 cells,Th17 cells,and Treg cells were analyzed by flow cytometry,enzyme linked immunosorbent analysis (ELISA) was used to detect the levels of interleukin (IL)-9,IL-10,IL-17A and transforming growth factor (TGF)-β1 in serum and bronchoalveolar lavage fluid (BALF) ,and real-time quantitative polymerase chain reaction (PCR) to detect the mRNA expression levels of IL-9,IL-10,IL-17A and TGF-β1 in lung tissues,Immunohistochemistry and Western Blot methods were used to detect the protein levels of PU.1,retinoic acid-related orphan receptor γt(RORγt) and forkhead transcription factor 3 (Foxp3) protein in lung tissues. Results Compared with the model group,the asthma symptoms in mice in hyperoside high-dose group were reduced,Ri and Re were significantly reduced,Cldyn was significantly elevated,the counts of white blood cells and eosinophils (EOS) in BALF were significantly reduced,the proportions of Th9 and Th17 cells were reduced,and the proportion of Treg cells was elevated,and the mRNA levels of IL-9 and IL-17A in the lung tissues as well as those in serum and BALF were significantly reduced,and levels of IL-10 and TGF-β1 in lung tissues as well as those in serum and BALF were significantly increased, the protein expression levels of PU. 1 and RORγt in lung tissues were significantly reduced,and protein expression level of Foxp3 in lung tissues was significantly increased,the differences being all statistically significant (P<0.05) . Conclusion Hyperoside can inhibit Th9,Th17 cell differentiation and Treg cell defferentiation as well as reducing airway inflammation in allergic asthma mice.
[中图分类号]
R285.5
[基金项目]
吉林省科技发展计划项目 (编号:20200404043YY)