盐酸氨溴索联合布地奈德对致敏大鼠气道炎症及肺部氧化应激的影响

Effect of ambroxol hydrochloride combined with budesonide on the airway inflammation and lung oxidative stress in allergized rats

目的研究盐酸氨溴索与布地奈德对致敏大鼠气道炎性反应及肺部氧化应激水平的影响。方法将40只健康清洁级SD雄性大鼠随机分为空白对照组、模型组、阳性对照组和实验组,每组各10只。除空白对照组外,其余各组大鼠均以卵白蛋白致敏,构建大鼠哮喘模型。模型组分别于第1,8天腹腔注射10%卵白蛋白1 mL,第15天后雾化吸入1%卵白蛋白,每天1次,每次0.5 h,至大鼠出现腹肌收缩、点头等;阳性对照组每次雾化前1 h雾化吸入0.02%布地奈德30 min;实验组在阳性对照组的基础上,吸入布地奈德前25 min腹腔注射盐酸氨溴索60 mg·kg^(-1)。各组均干预10周。用细胞计数法检测各组大鼠肺泡灌洗液中淋巴细胞及嗜酸性粒细胞比例;用苏木精-伊红(HE)染色法观察各组大鼠气道组织病理学变化;用比色法检测各组肺组织中氧化应激指标水平。结果空白对照组、模型组、阳性对照组及实验组的嗜酸性粒细胞比例分别为(1.29±0.41)%,(81.44±5.15)%,(37.51±5.38)%,(20.14±4.65)%,淋巴细胞比例分别为(4.01±0.87)%,(17.38±2.99)%,(11.59±2.01)%,(7.96±1.43)%,实验组与模型组及阳性对照组比较,差异均有统计学意义(均P<0.01)。HE染色显示,模型组病理变化最为明显,阳性对照组次之。空白对照组、模型组、阳性对照组及实验组大鼠肺组织中谷胱甘肽(GSH)水平分别为(1.54±0.15),(0.53±0.18),(0.79±0.09),(1.15±0.11)μmol·L^(-1);活性氧(ROS)水平分别为(1643±163),(4986±195),(3619±184),(2348±171)U·mg^(-1)pro。实验组GSH、ROS与阳性对照组相比,差异均有统计学意义(均P<0.01)。结论盐酸氨溴索与布地奈德联合可有效减轻致敏大鼠气道炎性反应及肺组织氧化应激损伤,可有效治疗哮喘急性发作。

Objective To explore the effect of ambroxol hydrochloride combined with budesonide on the airway inflammation and lung oxidative stress in allergized rats. Methods Forty cases of clean health level SD male rats were randomly divided into blank control group,model group,positive control group and test group,10 rats in each group. Except blank control group,the rats of other groups were treated with ovalbumin to build the asthma model. Model group was intraperitoneally injected 10% ovalbumin 1 mL on 1,8 d,and 1% ovalbumin was inhaled on 15 d,once a day for 0. 5 h,till the rats appear abdominal muscle contraction,nodding and so on. Positive control group was inhaled 0. 02%budesonide 30 min before nebulization for 1 h before nebulization. The test group was intraperitoneal injected ambroxol hydrochloride 60 mg·kg^-1 at 25 min before inhalation of budesonide on the basis of positive control group. Each group were intervention for 10 weeks. The numbers of lymphocytes and eosinophils in bronchoalveolar lavage fluid of rats in each group were measured by cell counting method. The airway histopathological and morphological changes were observed by hematoxylin-eosin（ HE） staining. Colorimetric method was used to detect the levels of oxidative stress in lungs of each group. Results The percentages of eosinophils in blank control group,model group,positive control group and test group were（ 1. 29 ± 0. 41） %,（ 81. 44 ± 5. 15） %,（ 37. 51 ± 5. 38） %,（ 20. 14 ± 4. 65） %,the percentages of lymphocytes were（ 4. 01 ± 0. 87） %,（ 17. 38 ± 2. 99） %,（ 11. 59 ± 2. 01） %,（ 7. 96 ± 1. 43） %. The difference betweent test group and model group and positive control group was statistically significant（ P〈0. 01）. HE staining showed that the most obvious pathological changes in model group,followed by positive control group. The levels of glutathione（ GSH） in lung tissue of blank control group,model group,positive control group and test group were（ 1. 54 ± 0. 15）,（ 0. 53 ± 0. 18）,（ 0. 79 ± 0. 09）,（ 1. 15 ± 0. 11） μmol · L^-1,and reactive oxygen species（ ROS） were（ 1643 ± 163）,（ 4986 ± 195）,（ 3619 ± 184） and（ 2348 ± 171） U · mg～（-1） pro. The GSH,ROS in test group and positive control group had significant difference（ P〈0. 01）. Conclusion Ambroxol hydrochloride combined with budesonide can relieve the airway inflammation and lung oxidative stress in rats with asthma,which can be used for the treatment of acute asthma attacks.