The Influence And Mechanism Of Budesonide Combined With Montelukast To Airway Remodeling Of Asthma Mice

Objective:To investigate the influence of budesonide combined with montelukast to airway remodeling on the asthma model of mouse and the role of OPN (osteopontin, OPN) and transforming growth factor (31 (Transforming Growth Factor-(31, TGF-β1) in asthma airway remodeling in mice.Methods:Forty 6 to 8 weeks healthy male BALB/c mice were randomly divided into five groups, eight mice in each group:(1) Control group (CON group), (2) Airway remodeling asthma model (AM group), (3) Budesonide group (BUD group), (4) Montelukast group (MK group), (5) Budesonide and montelukast group (BUD-MK group). AM group mice were sensitized by intraperitoneal injection of ovalbumin(OVA) solution adsorbed with aluminum hydroxide and were challenged by OVA solution. CON group mice were treated with normal saline instead at the same dose. BUD group mice were treated with budesonide(1 mg per each time) after sensitized. MK group mice were treated with MK (10mg per kg weight) after sensitized, and BUD-MK group mice were treated with budesonide combined with MK after sensitized. Mice were killed after 24 hours of the last challenge. The numbers of eosinophile in the bronchoalveolar lavage fluid (BALF) were counted. The lung tissue of mice was removed for histological examination. Lung tissues were sliced and stained with HE. The expression of OPN in airway tissues was observed by immunohistochemistry. The Masson staining was used to observe collagen deposition on airway. The content of OPN and TGF-β1 in serum was measured by ELISA.Results:(1) The counts of eosinophile in the BALF:Compared with CON group (4.12±1.35±106/L), the number of EOS in AM group (22.3±1.95×106/L) was significantly increased (P<0.05). BUD group (14.57±1.62×106/L) and MK group(16.89±1.34×106/L) were significantly less than AM group (P<0.05), BUD-MK group (10.43±1.13×106/L) was not much less than BUD group (P>0.05). (2) The pathological changes of lung tissue:Compared with CON group, the infiltration of eosinophils, lymphocytes and other inflammatory cells was observed in lung tissue of AM group, which was worse than in BUD group and BUD-MK group. (3) The expression of OPN in lung tissue: Compared with CON group, more positive cells were observed in AM group. But, the positive cells in BUD group, MK group and BUD+MK group were less than in AM group. (3) The contents of cytokines in the BALF:①OPN:Compared with CON group (1147.59±225.66pg/ml), the contents of OPN in BALF was significantly increased (P<0.05) in AM group (4222.37±707.28pg/ml). But, in BUD group (1762.37±354.47pg/ml) and MK group (2150.46±423.77pg/ml) were lesser than in AM group. The contents of OPN in BALF was decreased significantly in BUD+MK group (1433.912±235.67) compared with that in AM group, BUD group and MK group (P＜0.05).②TGF-β1:Compared with CON group (30.63±5.17pg/ml), the contents of TGF-β1 in BALF was significantly increased(P＜0.05) in AM group(102.83±21.77pg/ml). The contents of TGF-β1 in BALF was decreased significantly in BUD group (68.32±20.98pg/ml) and MK group (60.21±14.20) than in AM group (P＜0.05). The contents of TGF-β1 in BUD-MK (44.12±10.12pg/ml) group was decreased significantly than in BUD group and MK group (P＜0.05).③There was linear positive correlation between the contents of OPN and TGF-beta 1 in AM group(r= 0.927,P＜0.01). (3) Masson staining:①The quantity of collagen deposition was increased significantly in AM group (5.12±0.42) than in CON group (1.68±0.28), (P＜0.05). The quantity of collagen deposition was decreased significantly in BUD group (2.94±0.32), MK group (3.05±0.35) and BUD+Mk group (2.48±0.25) than in AM group(P＜0.05). Compared with BUD+MK group, BUD group and MK group were significantly increased(P＜0.05).②There was a linear positive correlation between the contents of OPN and quantity of collagen deposition in AM group (r=0.929, P＜0.01).Conclusions:1. After inhaled ovalbumin two weeks, asthmatic models of airway remodeling were established in mice.2. TGF-β1 and OPN taken part in the process of airway remodeling of asthma, and there was a linear positive correlation between them.3. BUD and MK maybe inhibit the airway remodeling by reducing the expression of TGF-β1 and OPN.4. Budesonide combined with Montelukast is a better curative effect than treatment of using budesonide or Montelukast alone on the airway remodeling of asthma.