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血管活性肠肽对哮喘小鼠气道炎症及Th17/Treg平衡的影响
Effects of vasoactive intestinal peptide on airway inflammation and Th17/Treg balance in asthmatic mice
目的 探讨血管活性肠肽（VIP）对哮喘小鼠气道炎症的影响，以及对Th17细胞/调节性T细胞（Treg）失衡的调控作用。方法 将30只BALB/c小鼠随机分成对照组、哮喘组、VIP组，每组10只。利用卵白蛋白（OVA）致敏和激发制作急性哮喘小鼠模型；对照组致敏和激发阶段均以生理盐水代替OVA；VIP组每次OVA激发前，先以VIP溶液（20 μg/mL）雾化吸入30 min。留取各组小鼠支气管肺泡灌洗液、肺组织等标本。利用苏木精-伊红染色观察肺组织病理变化；ELISA法检测肺泡灌洗液中Th17/Treg相关细胞因子水平；免疫组化及Real-time PCR检测肺组织中Th17细胞特异性转录因子RORγt及Treg特异性转录因子Foxp3表达情况。结果 病理组织学结果显示VIP组小鼠肺组织气道炎症表现较哮喘组减轻。哮喘组小鼠BALF中IL-17浓度高于对照组（P < 0.01），VIP组IL-17浓度较哮喘组降低（P < 0.01），但仍高于对照组（P < 0.01）。哮喘组BALF中IL-10浓度低于对照组（P < 0.01），VIP组IL-10浓度较哮喘组升高（P < 0.01），但仍低于对照组（P < 0.01）。哮喘组小鼠肺组织RORγt mRNA及蛋白表达高于对照组（P < 0.01），Foxp3 mRNA及蛋白表达低于对照组（P < 0.01）；VIP组肺组织RORγt mRNA及蛋白表达水平低于哮喘组（P < 0.01），Foxp3 mRNA及蛋白表达水平高于哮喘组（P < 0.05）。结论 哮喘小鼠存在Th17/Treg免疫失衡，VIP可通过调控Th17/Treg免疫失衡而改善哮喘小鼠气道炎症。
Objective To investigate the effects of vasoactive intestinal peptide (VIP) on the airway inflammation and its regulatory effect on Th17/Treg imbalance in asthmatic mice. Methods A total of 30 BALB/c mice were equally and randomly divided into three groups:control, asthma, and VIP. An acute asthmatic mouse model was established by sensitization and challenge with ovalbumin (OVA). The control group received normal saline instead of OVA. Before the challenge with OVA, the VIP group was administered VIP (20 μg/mL) by aerosol inhalation for 30 minutes. The bronchoalveolar lavage fluid (BALF) and the lung tissue were collected from mice. The pathological changes in the lung tissue were observed by hematoxylin and eosin staining. The levels of Th17/Treg-related cytokines in BALF were measured by enzyme-linked immunosorbent assay. The expression of retinoid-related orphan receptor gamma t (RORγt) and forkhead box P3 (Foxp3) were measured by real-time fluorescence quantitative PCR and immunohistochemistry. Results The histopathological results showed that the VIP group had milder symptoms of airway inflammation than the asthma group. The level of IL-17 in BALF in the asthma group was significantly higher than that in the control group and the VIP group (P < 0.01), but the level of IL-17 in the control group was significantly lower than that in the VIP group (P < 0.01). The level of IL-10 in BALF in the asthma group was significantly lower than that in the control group and the VIP group (P < 0.01, but the level of IL-10 in the VIP group was significantly higher than that in the control group (P < 0.01). The asthma group showed significantly higher expression levels of RORγt mRNA and protein in the lung tissue and significantly lower expression levels of Foxp3 mRNA and protein than the control group (P < 0.01). The VIP group had significantly lower expression levels of RORγt mRNA and protein in the lung tissue and significantly higher expression levels of Foxp3 mRNA and protein than the asthma group (P < 0.05). Conclusions The Th17/Treg imbalance may be closely related to the airway inflammation in asthmatic mice. VIP can improve airway inflammation by regulating the Th17/Treg imbalance in asthmatic mice.
血管活性肠肽 / 哮喘 / Th17细胞 / 调节性T细胞 / 小鼠
Vasoactive intestinal peptide / Asthma / Th17 cell / Regulatory T cell / Mice
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