| 摘要: |
| 目的探讨雷帕霉素对哮喘缓解期小鼠气道炎症的作用与机制。方法将50只Balb/c小鼠随机分为5组：正常对照组、哮喘缓解组、哮喘模型组、地塞米松组和雷帕霉素组，每组10只。采用卵蛋白致敏与激发制备哮喘模型，休息3周后，采用卵蛋白再次激发1周，在小鼠休息期间治疗组采用雷帕霉素或地塞米松干预。采用Buxco无创肺功能仪检测气道高反应性；Luminex测定肺泡灌洗液IL-4、IL-5、IL-13、IL-17及INF-γ水平；肺组织HE染色评价观察小鼠肺组织气道炎症；Westernblot检测肺组织p-S6、S6、AKT、p-AKT（S473）蛋白含量；流式分析测定肺泡灌洗液CD4+CD25+Foxp3+Treg细胞百分率。结果与正常组及哮喘缓解组比较：哮喘模型组气道反应性升高（P＜0.01），灌洗液IL-4、IL-5、IL-13及IL-17水平增加（均P＜0.01），肺组织气道炎症评分增高（P＜0.01），肺组织mTORC1信号通路下游蛋白p-S6蛋白增加（P＜0.01），灌洗液Treg细胞比例上升（P＜0.01）；与模型组比较：雷帕霉素在乙酰甲胆碱浓度为6.25mg/ml时,可降低Penh值（P＜0.01），同时降低肺泡灌洗液IL-4水平（P＜0.01），并抑制肺组织炎症细胞浸润（P＜0.01），而肺组织mTORC1信号通路下游蛋白p-S6蛋白下降（P＜0.01），但雷帕霉素可降低肺泡灌洗液Treg细胞比例（P＜0.01）。结论在哮喘缓解期应用雷帕霉素，其可通过mTORC1信号通路显著抑制哮喘急性发作时气道炎症；在哮喘缓解期应用雷帕霉素，对控制哮喘再次发作具有积极作用。 |
| 关键词: 雷帕霉素 哮喘 mTOR 信号通路 |
| DOI：10.12056/j.issn.1006-2785.2017.39.11.2017-19 |
| 分类号: |
| 基金项目:国家自然科学基金（81403247/H2902）；浙江省医药卫生科技项目（2014KYA170） |
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| Effects of rapamycin on airway inflammation in mice with asthma during remission |
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JIN Hualiang, WANG Limin, WANG Jiaoli, Xiajunbo, MA Shenglin
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Hangzhou First People's Hospital/Hangzhou Hospital of Nanjing Medical University
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| Abstract: |
| Objective To investigate the effects of rapamycin on airway inflammation in mice with asthma during remission. Methods Fifty Balb/c mice were randomly divided into 5 groups: normal control, asthma, asthma in remission, dexamethasone and rapamycin groups with 10 mice in each group. The asthma model was induced by ovalbumin (OVA) sensitization and challenge; after 3-week interval the mice were challenged by OVA again. Dexamethasone, rapamycin or saline was given to 3 groups, respectively during 3-week interval. Airway reactivity was measured by Buxco's non-invasive system. Cytokines IL-4, IL-5, IL-13, IL-17 and INF-γ in bronchoalveolar lavage fluid (BALF) were assessed by Luminex method. Lung tissues were examined for inflammatory cell infiltration. The expression of p-S6, S6, AKT and p-AKT (S473) in lung tissue was examined by Western blot. CD4+CD25+Foxp3+ Treg cells in BALF was assessed by flow cytometry. Results Compared with the
normal and remission groups, OVA re-expose significantly increased airway hyperresponsiveness(P＜0.01), elevated IL-4, IL-5, IL-13, IL-17 levels (P＜0.01), and reduced INF-γ in the BALF (P＜0.01); it also markedly increased inflammatory cells in lung tissue, increased Ttreg cells in BALF and decreased expression of p-S6 (P ＜0.01). Rapamycin significantly reduced airway hyperresponsiveness to aerosolized methacholine at 6.25mg/ml(P＜0.01), inhibited IL-4 level in BALF, and markedly decreased inflammatory infiltration in lung tissues(P＜0.01). Notably, rapamycin significantly inhibited the expression of p-S6; and also Treg cells in BALF were significantly reduced after rapamycin treatment. Conclusion Antiasthmatic effects of rapamycin during re- mission time are at least partially through mTORC1 signaling pathway, and rapamycin may be used for asthma patients in re-
mission to control asthma attack. |
| Key words: Rapamycin Asthma mTOR signaling pathway |
