Expression of Notch ligands and the effects of Budesonide in airway remodeling in mice with asthma
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摘要:
目的探讨哮喘小鼠气道重塑发病过程中Notch配体Jagged1及Jagged2在肺组织的表达变化及布地奈德的干预作用。 方法清洁级C57BL/6雄性小鼠30只随机分为对照组、哮喘气道重塑组、布地奈德干预组，每组各10只。建立哮喘气道重塑模型，利用酶联免疫吸附测定、免疫组化、Western blot等方法，检测炎症因子、气道重构和Jagged1及Jagged2表达变化，并分析它们的相关性。 结果哮喘气道重塑组小鼠肺组织的炎症因子IL-4/5/13、Jagged1及Jagged2表达明显增加（P < 0.01或0.001）；布地奈德干预后，气道炎症、气道粘液分泌及胶原沉积明显缓解（P < 0.01），Jagged1及Jagged2表达明显降低（P < 0.05或0.01）；Jagged1及Jagged2表达水平与炎症因子、气道重构呈正相关（P < 0.001）。 结论Notch配体Jagged1及Jagged2表达增加可能参与了哮喘小鼠气道炎症反应及气道重塑。布地奈德能抑制哮喘气道炎症反应及气道重塑可能与降低Jagged1及Jagged2的表达有关。 Abstract:ObjectiveTo investigate expression of Notch ligands (Jagged 1 and Jagged 2) and the effects of Budesonide in airway remodeling in mice with asthma. MethodsThirty C57BL/6 male mice were randomly divided into 3 groups (10 mice/group) including control group, asthma group, and Budesonide group. The pathological changes of the airway were assessed by hematoxylin and eosin staining. The cytokines in the lung homogenate were assessed by ELISA. The immunohistochemistry and western blot was used to estimate the expression of Jagged1 and Jagged2 in the lung tissues. The correlations among them were also analyzed. ResultsCompared with the control group, the expressions of IL-4/5/13, Jagged1 and Jagged2 in the lung tissues were significantly increased (P < 0.01 or 0.001), but decreased significantly in the Budesonide-treated asthma group. Both Jagged1 and Jagged2 were positively correlated with the cytokines, mucus production and collagen deposition (P < 0.001). ConclusionJagged 1 and Jagged 2 may be associated with airway remodeling in mice with asthma. Budesonide can improve airway remodeling, possibly by decreasing the expression of Jagged1 and Jagged2. -
Key words:
- asthma /
- airway remodeling /
- Notch pathway /
- budesonide
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图 1 布地奈德明显缓解了哮喘小鼠气道炎症
A:肺泡灌洗液中细胞总数; B:肺泡灌洗液中嗜酸性粒细胞分类计数; C-E:肺组织HE染色(箭头所示, 原放大倍数: × 200); F: HE评分半定量评估气道炎症浸润, **P < 0.01; ***P < 0.001.
图 2 布地奈德缓解气道炎症反应
A:肺组织匀浆IL-4的表达水平; B:肺组织匀浆IL-5的表达水平; C:肺组织匀浆IL-13的表达水平水平的表达. *P < 0.05; **P < 0.01; ***P < 0.001.
图 3 布地奈德缓解哮喘小鼠的胶原沉积及粘液高分泌
A-C:为肺组织Masson染色(箭头所示，×200倍); D:为Masson染色的半定量评分; E-G:为肺组织PAS染色(箭头所示, 原放大倍数: ×200); H:为PAS染色的半定量评分. *P < 0.05, **P < 0.01, ***P < 0.001.
图 4 布地奈德缓解哮喘小鼠肺组织的Jagged1及Jagged2
A-C, E-G:用免疫组化方法检测各组小鼠肺组织的Jagged1及Jagged2表达（原放大倍数: ×400）; I:为进一步利用Western Blot方法检测肺组织的Jagged1及Jagged2的蛋白表达水平; D, H, G, K:为利用Image-pro Plus图像分析软件进行半定量分析Jagged1及Jagged2表达水平. *P < 0.05; **P < 0.01; ***P < 0.001.
表 1 Jagged1及Jagged2表达水平与炎症因子、气道重构的相关性分析
参数 Jagged1 Jagged2 r P值 r P值 IL-4 0.75 0.0012 0.6 0.0188 IL-5 0.87 < 0.0001 0.72 0.0023 IL-13 0.73 0.0019 0.55 0.0349 HE Scores 0.77 0.0007 0.72 0.0027 Masson Scores 0.86 < 0.0001 0.83 0.0001 PAS Scores 0.8 0.0004 0.81 0.0003 
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