吸入布地奈德对烟熏诱导小鼠气道重构的影响

Effects of inhaled budesonide on airway remodeling driving by cigarette smoke in mice

目的观察吸入布地奈德对烟熏诱导小鼠气道重构的干预作用,并比较早、晚期治疗的差异。方法 (1)60只C57BL/6小鼠分为4组,对照组共20只,其中5只于第5~8周予生理盐水5 ml雾化吸入30 min,5只于9~12周予生理盐水5 ml雾化吸入30 min,2次/d。模型组共20只,全部予烟熏处理8周,1 h/次,2次/d,其中5只于第5~8周予生理盐水5 ml雾化吸入30 min,5只于9~12周予生理盐水5 ml雾化吸入30 min,2次/d。早期治疗组共10只,全部予烟熏处理8周,于第5~8周每次烟熏1 h前予布地奈德混悬液雾化吸入30 min。晚期治疗组:共10只,全部予烟熏处理8周,于第9~12周每次烟熏1h前予布地奈德雾化吸入30 min。(2)于第4周结束处死对照组、模型组各10只、12周结束处死其余全部小鼠,取肺组织分别行HE和Masson染色、免疫组化检测转化生长因子β1(TGF-β1)表达以及高效液相法测定胫脯氨酸(HYP)含量。图像分析软件测定HE染色下基底膜周径(Pbm)、支气管管壁总面积(WAt)、Masson染色下支气管壁胶原面积(Wcol)。结果早期治疗组的气道重构程度及TGF-β1表达均低于模型组(12周)及晚期治疗组,但较对照组、模型组(4周)增加(P<0.05)。晚期治疗组气道重构程度及TGF-β1表达与模型组(12周)无显著差异(P>0.05)。结论早期吸入布地奈德能延缓但并不能逆转烟熏诱导气道重构的发生,其机制可能与降低TGF-β1表达相关。

Objective To observe the intervention of inhaled budesonide on airway remodeling driving by cigarette smoke in mice,and find out the therapeutic differences between early phase and late phase. Methods ①60 C57BL/6 mice were divided into four groups, including control group （n=20）, model group （n=20）, early stage treatment group （n=10） and late stage treatment group （n=10）. During week 5 to week 8, atomization of normal saline was given to 5 mice for 30 minutes （twice/day） in the control group and the model group. During week 9 to week 12, the same treatment was given to other 5 mice in the control group and the model group. All mice in the model group, early stage treatment group and late stage treatment group were exposed to cigarette smoke for 8 weeks （1 hour each time,twice/day）. In the early stage treatment group, 1 mg/5 ml budesonide was inhaled at one hour before cigarette smoke explosion during week 5 week 8.The same treatment was given to the late stage treatment group during week 9 to week 12. ②Termination of 10 mice took place in control group and model group at the end of week 4. Other all mice were terminated at the end of week 12. The lung tissues were moved used for hematoxylin-eosin, Masson and immunohistochemical stainings as well as the measurement of hydrocarbon proline content.The length of basement membrane of the epithelium （Pbm）, the area of the brochial airway wall （Wat） and the are of the collagen （Wcol） were assessed by Image-Pro Plus. Resluts The airway remodeling degree and the TGF-β1 expression were weaker in the early treatment stage group than the model group （week 12） and the late treatment stage group （ P 〈0.05）, but higher than the control group and the model group （week 4）. There were no differences between the model group （week 12） and the late treatmen stage group （ P 〉0.05）. Conclusion The early treatment of inhaled budesonide can delay but not reverse the airway remodeling driving by cigarette smoke. And its mechanism is possibly connected with reducing the expression of TGF-β1.