盐酸氨溴索对毛细支气管炎小鼠IL-17/STAT3通路及气道高反应的影响

Effects of ambroxol hydrochloride on IL-17/STAT3 pathway and airway hyperresponsiveness in mice with bronchiolitis

目的探究盐酸氨溴索对毛细支气管炎小鼠气道高反应性及白细胞介素17(IL-17)/信号传导转录激活因子3(STAT3)通路的影响。方法采用随机数字表法将72只Balb/c小鼠随机分为对照组、模型组、布地奈德组(20 mg/kg)、盐酸氨溴索高、中、低剂量(60、30、15mg/kg)组,每组12只。除对照组外,其余各组小鼠用呼吸道合胞病毒(RSV)混悬液滴鼻法复制毛细支气管炎小鼠模型。造模完成后,给药组小鼠雾化吸入相应剂量的药物,对照组和模型组小鼠雾化吸入生理盐水,1次/d,连续7 d。双腔体描计法检测小鼠清醒状态下的小鼠气道阻力(sRaw);检测血清中IL-17、IL-6、IL-23水平;空斑法测定肺组织匀浆中病毒滴度;苏木精-伊红染色(HE)观察肺组织病理变化;蛋白免疫印迹法(Western blotting)检测肺组织中IL-17、p-STAT3、STAT3蛋白的表达。结果与对照组相比,模型组小鼠s Raw,血清IL-17、IL-6、IL-23水平,肺组织匀浆病毒滴度,IL-17、p-STAT3/STAT3蛋白表达显著升高(P<0.05),肺部支气管及肺泡结构明显破坏,大量炎性细胞浸润,支气管气道变窄;与模型组相比,布地奈德组和盐酸氨溴索高、中剂量组小鼠s Raw,血清IL-17、IL-6、IL-23水平,肺组织匀浆病毒滴度,IL-17、p-STAT3/STAT3蛋白表达明显降低(P<0.05),肺组织炎症明显减轻。结论盐酸氨溴索可抑制炎性因子的表达,减轻毛细支气管炎小鼠气道炎症反应和气道高反应性;其作用机制可能与阻断IL-17/STAT3途径有关。

Objective To investigate the effects of ambroxol hydrochloride on airway hyperresponsiveness and interleukin-17(IL-17)/signal transduction activator of transcription 3(STAT3)pathway in mice with bronchiolitis.Methods Seventy-two Balb/c mice were randomly divided into control group,model group,budesonide group(20 mg/kg),high,medium and low dose ambroxol hydrochloride groups(60,30,15 mg/kg)according to the random number table,with twelve mice in each group.Except for the control group.The mice in other groups were made into bronchiolitis model by nasal drip of respiratory syncytial virus(RSV)suspension.After modeling,mice in the drug groups were given the corresponding dose of drugs,while mice in the control group and model group were inhaled with normal saline,once a day,for 7 d.The airway resistance(sRaw)in conscious mice was measured by dual chamber plethysmography.The levels of serum IL-17,IL-6,and IL-23 were detected.The virus titer in lung homogenate were measured by plaque method.The pathological changes of lung tissue were observed by HE staining,the expression of IL-17,p-STAT3,and STAT3 in lung tissue were detected by Western blotting.Results Compared with those in the control group,sRaw,levels of serum IL-17,IL-6 and IL-23,virus titer in lung homogenate,protein expression of IL-17 and p-STAT3/STAT3 of mice in model group were significantly higher(P<0.05),the structure of bronchus and alveoli of lung were destroyed,a large number of inflammatory cells infiltrated,and the bronchial airway became narrow.Compared with those in the model group,sRaw,the levels of serum IL-17,IL-6,and IL-23,virus titer in lung homogenate,protein expression of IL-17 and p-STAT3/STAT3 of mice in budesonide group,high and medium dose ambroxol hydrochloride groups were significantly lower(P<0.05),the inflammation of lung tissue was significantly reduced.Conclusion Ambroxol hydrochloride can inhibit the expression of inflammatory factors,reduce airway inflammation and airway hyperresponsiveness in mice with bronchiolitis,and its mechanism may be related to blocking IL-17/STAT3 pathway.