布地奈德干预对卵白蛋白致敏小鼠抗原激发后气道炎症及气道重塑的影响

Effects of budesonide on airway inflammation and airway remodeling in the ovalbumin sensitized and challenged mice

目的 观察早期及延迟应用布地奈德对支气管哮喘 (简称哮喘 )小鼠气道炎症和气道重塑的防治作用。方法 40只小鼠分为 5组,每组 8只。鸡卵白蛋白(OVA)致敏 /激发组 (A组 ),生理盐水对照组(B组),布地奈德 (BUD)早期治疗组 (C组 ),BUD延迟治疗组 (D组 ),OVA延迟对照组(E组)。小鼠于第 0、14天以OVA致敏,从第 1次致敏后第 24天开始雾化吸入 2.5%的OVA激发并持续 18d,建立气道重塑模型;分别在早期(抗原激发前 1d始)和延迟(第 1次抗原激发后第 18天始)雾化吸入BUD(0.5mg/ml),观察抗原激发及BUD应用后支气管肺泡灌洗液(BALF)中嗜酸粒细胞(EOS)数、上清液白细胞介素 5(IL-5)和γ干扰素 (IFN-γ)水平的变化,同时对肺组织切片行苏木精 伊红(HE)、过碘酸雪夫(PAS)、Masson三色染色,测定支气管壁周围EOS计数、定量杯状细胞百分比及黏液分泌评分并测定气道平滑肌增生高度及胶原面积。结果 经过反复抗原激发,A组BALF中EOS数为(57.460±11 060)×104 /ml,B组为[ (0.050±0.020)×104 /ml],两组比较差异有统计学意义(P<0.01);A组BALF中IL- 5水平及IFN -γ水平分别为(52.9±2.8)pg/ml、(39.5±3.2)pg/ml,B组分别为(16.8±1.5)pg/ml、(63.8±3.3)pg/ml,两组比较差异有统计学意义 (P<0.01 );A、B组支气管壁周围EOS数分别?

Objective To investigate the effects of budesonide(BUD) used in an early phase or delayed phase on the airway inflammation and airway remodeling in ovalbumin(OVA) sensitized and challenged mice. Methods Forty mice were divided into 5 groups(8 in each group) including group A[ovalbumin(OVA) sensitized/challenged mice],group B(saline sensitized/challenged mice),group C(OVA sensitized/challenged mice treated by BUD in the early phase),group D(OVA-sensitized/challenged mice treated by BUD in the late phase) and group E(OVA-sensitized/challenged mice following saline challenge for 18 days). Mice were sensitized on days 0 and 14 by OVA and challenged from days 24 to 41 by OVA repeatedly to establish a murine model of asthma characterized by airway inflammation and airway remodeling. To assess the effects of BUD on the development of airway remodeling and on established airway remodeling,animals were treated with aerosolized BUD(0.5 mg/ml per day) from day 1 before OVA challenge(group C) and from day 18 following first OVA challenge(group D). The outcome measurements included airway inflammatory indices,eosinophils(EOS) count and amount of collagen deposition around the bronchus,area of airway smooth muscle,the degree of mucus secretion in the lumens and depth of airway smooth muscle(ASM) in different grade bronchus by HE,PAS and Masson′s staining. The bronchoalveolar lavage fluids(BALF) were assayed for IL-5 and IFN-γ levels. Results In repeatedly OVA-challenged mice(group A),EOS counts in BALF increased significantly[(57.460±11.060)×10~4/ml] when compared with group B[(0.050±0.020)×10~4/ml,P<0.01]. The IL-5 level increased significantly[(52.9±2.8)(pg/ml vs) (16.8±1.5)pg/ml,P<0.01] and IFN-γ decreased significantly[(39.5±3.2)pg/ml vs ((63.8±3.3))pg/ml,P<0.01]. Repeatedly OVA-challenged animals(group A) also developed an increase in EOS counts around bronchus[(1 018±118)/mm^2] when compared with group B[(7±3)/mm^2,P<0.01],goblet cell hyperplasia[(46.0±5.8)% vs (1.8±0.5)%,P<0.01]and mucus hypersecretion[(score 2.98±0.23) vs (score 0.13±0.06),P<0.01],airway smooth muscle hypertrophy[(30.2±2.2)/μm^2/μm vs (13.1±1.0)μm^2/μm,P<0.01],enhanced collagen deposition of reticular basement membrane(RBM)[(24.9±1.3)μm^2/μm vs (4.3±0.6)μm^2/μm,all P<0.01]. Early treatment with BUD significantly reduced EOS counts in BALF [(7.140±1.250)×10~4/ml] as compared with group A[(57.460±11.060)×10~4/ml],P<0.01]. Early BUD treatment also significantly reduced EOS counts around bronchus [(214±26)/mm^2],allergen-induced structural changes including goblet cell hyperplasia[(16.1±2.5)%] and mucus hypersecretion(1.10±0.15),airway smooth muscle hypertrophy[(14.0±0.7)μm^2/μm],and RBM collagen deposition[(12.6±1.3)μm^2/μm]. In group E,EOS counts in BALF[(1.250±0.330)×10~4/ml] were decreased significantly when compared with group A(P<0.01),but airway smooth muscle hypertrophy[(32.4±1.8)μm^2/μm],and RBM collagen deposition[(22.8±1.7)μm^2/μm] showed no reduction as compared with group A( all P>0.05). Delayed BUD treatment significantly reduced EOS counts in BALF[(0.800±0.170)×10~4/ml],goblet cell hyperplasia [(29.3±4.3)%] and mucus hypersecretion(1.63±0.17,P<0.05 or P<0.01),but had no effects on OVA-induced airway smooth muscle hypertrophy[(30.1±1.8)μm^2/μm] and RBM collagen deposition[(23.7±1.4)μm^2/μm] as compared with group A(P>0.05). Conclusions Early treatment with BUD inhibited the development of airway inflammation of airway remodeling. However,delayed use with BUD inhibited airway inflammation,but only partially reversed airway remodeling in a murine model of asthma.