目的 研究布地奈德(BUD)对慢性哮喘小鼠支气管上皮细胞的血清类黏蛋白1样蛋白3(ORMDL3),基质金属蛋白酶-9(MMP-9)表达的影响,及其对气道重塑的干预作用。方法 应用BALB/C小鼠随机分为对照组、哮喘组、BUD组,用OVA(卵清蛋白)和氢氧化铝建立哮喘小鼠模型,为进一步了解布地奈德对气道重塑的干预机制,第21天BUD组小鼠在OVA激发前30 min吸入布地奈德。12周后,用苏木精-伊红染色(HE staining)分析气道壁厚度改变,Masson染色观察气道胶原沉积,通过免疫组织化学染色法(immunohistochemistry staining),蛋白质印迹法(western blot),实时荧光定量PCR(real-time PCR)测定小鼠支气管上皮细胞ORMDL3,MMP-9的表达含量。结果 12周后,哮喘组和BUD组的气道壁厚度增加,胶原蛋白沉积,ORMDL3,MMP-9表达明显高于对照组,BUD组气道病理改变程度和ORMDL3,MMP-9的升高幅度均不及哮喘组。经相关性分析可发现,MMP-9表达增加与ORMDL3基因的表达上调具有相关性,并且二者的表达增高与气道壁厚度有明显相关性。结论 布地奈德可能通过下调ORMDL3,MMP-9基因的表达,抑制慢性哮喘气道重塑进程。
Abstract
Objective To investigate the effect of budesonide on the expression of orosomucoid1-like3(ORMDL3),matrix metalloproteinases-9(MMP-9) and understand the mechanism of budesonide in inhibition of airway remodeling in chronic asthma mice. Method Mice were divided into three groups,including the asthmatic model group,the budesonide group and the control group.The asthmatic model were established by administrating OVA and aluminum hydroxide.The BUD group were administered with aerosol budesonide (100 μg/kg) before OVA challenge from day 21.The control group were sensitized and challenged with PBS instead.Using HE staining and Masson staining to assess the extent of bronchial inflammation and collagen deposition in each group.The expression of ORMDL3 and MMP-9 were examined in absence and presence of treatment via immunohistochemistry,RT-PCR and Western blotting in 12 weeks. Results The asthma group showed more inflammatory responses and airway remodeling compared with the control group and the BUD group.Administration of BUD decreased the severity of pathological changes but not eliminated these changes.Data proved that both ORMDL3 and MMP-9 increased significantly in asthma group while increased slightly in BUD group.Besides,the increase of ORMDL3 had a significant correlation with MMP-9 level.And content of ORMDL3,MMP-9 had positive correlation with the thickness of airway. Conclusion Administration of BUD may down-regulate the expression of ORMDL3 and MMP-9 to reduce airway remodeling.
关键词
气道重塑 /
慢性哮喘 /
布地奈德 /
血清类黏蛋白1样蛋白3 /
基质金属蛋白酶-9
Key words
budesonide /
orosomucoid 1-like 3 /
matrix metalloprpteinases-9 /
airway remolding /
chronic asthma
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基金
山东省自然科学基金项目(ZR2013HQ034)
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