布地奈德雾化吸入对哮喘小鼠肺泡灌洗液中炎性细胞分布和炎性因子表达水平的影响

Eeffect of nebulized budesonide inhalation on inflammatory cells distribution and expression level of inflammatory factors in bronchoalveolar lavage fluid in asthmatic mice

目的通过观察布地奈德雾化吸入治疗对哮喘小鼠肺泡灌洗液(bronchial alveolar lavage fluid,BALF)中炎性细胞分布和炎性因子白介素(interleukin,IL)-4、IL-5、IL-13及IL-17表达的影响,探索布地奈德治疗哮喘的可能作用机制。方法30只6~8周龄的雌性Balb/c小鼠按数字随机分为哮喘组、布地奈德治疗组和空白对照组,每组10只。卵蛋白腹腔注射和雾化激发构建哮喘模型,布地奈德组于雾化激发前给予布地奈德200 mg雾化吸入。于D21处死小鼠,收集BALF计数嗜酸性粒细胞和中性粒细胞比例,酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)法检测IL-4、IL-5、IL-13和IL-17的表达水平。结果成功构建哮喘小鼠模型。哮喘组小鼠BALF中嗜酸性粒细胞比例明显高于布地奈德组和空白对照组(P<0.017),布地奈德组嗜酸性粒细胞比例略高于空白对照组(P<0.017)。哮喘组小鼠BALF中中性粒细胞比例与布地奈德组相比,差异无统计学意义(P>0.017),但两组均明显低于空白对照组(P<0.017)。哮喘组小鼠BALF中IL-4、IL-5、IL-13和IL-17浓度均明显高于布地奈德组和空白对照组(P<0.017);并且布地奈德组IL-4、IL-5和IL-13浓度也明显高于空白对照组(P<0.017);但布地奈德组IL-17浓度与空白对照组相比,差异无统计学意义(P>0.017)。结论过敏性哮喘小鼠BALF以嗜酸性粒细胞增多为主。布地奈德能明显抑制嗜酸性粒细胞浸润和细胞因子IL-4、IL-5、IL-13和IL-17的表达,但对BALF中性粒细胞的浸润和分布影响较小。

Objective To explore the underlying mechanism of budesonide treating asthma,the inflammatory cells distribution and the expression of IL-4,IL-5,IL-13 and IL-17 in bronchoalveolar lavage fluid(BALF)of asthmatic mice treated by nebulized budesonide inhalation were observed.Methods Thirty 6-to-8-week-old female Balb/c mice were randomly divided into three groups:asthma group,budesonide treatment group and control group(10 mice in each group).Asthma models were established by ovalbumin(OVA)celiac injection and nebulized provocation.Mice in budesonide group inhaled 200 mg budesonide before nebulized provocation,and they were executed in D21.The data of eosinophil and neutrophil count and their percentage in BALF were collected,and the expression level of IL-4,IL-5,IL-13 and IL-17 in BALF was tested by enzyme-linked immunosorbent assay(ELISA).Results Asthma mice models were successfully established.Eosinophil percentage in asthma group was significantly higher than that in budesonide and control group(P<0.017),and that in budesonide group was significantly higher than that in control group(P<0.017).Neutrophil percentage in asthma group had no difference with that in budesonide group(P>0.017),but both were significantly lower than in control group(P<0.017).The expression of IL-4,IL-5,IL-13,and IL-17 in asthma group were significantly higher than those in budesonide and control group(P<0.017).The expression of IL-4,IL-5 and IL-13 in budesonide group were also significantly higher than those in control group,but the expression of IL-17 in budesonide group had no difference with that in control group(P>0.017).Conclusion The proliferation of eosinophil dominates BALF of mice with allergic asthma.Budesonide can suppress eosinophil infiltration and the expression of IL-4,IL-5,IL-13 and IL-17 well,but has poor effect on neutrophil infiltration and distribution in BALF.