小青龙汤对支气管哮喘大鼠黏液高分泌及气道炎症的影响

The Influence of Minor Green Dragon Decoction on Mucus Hypersecretion and Airway Inflammation in Rats with Bronchial Asthma

目的:观察小青龙汤对哮喘大鼠黏液高分泌及气道炎症的影响。方法:60只SPF级SD大鼠按随机数字表法分为对照组、模型组、氨溴索组及小青龙汤高剂量组(14 g·kg^(-1))、小青龙汤中剂量组(7 g·kg^(-1))、小青龙汤低剂量组(3.5 g·kg^(-1)),每组10只。除对照组外,其余组大鼠建立支气管哮喘模型。造模成功后,各组大鼠进行药物干预,对照组和模型组给予等体积生理盐水进行灌胃,每天1次,连续14天。大鼠肺功能仪检测大鼠第0.3秒用力呼气容积(forced expiratory volume,FEV0.3)、用力肺活量(forced vital capacity,FVC)、呼气峰流量(peak expiratory flow,PEF)及气道阻力值(air way resistance,R);瑞氏-姬姆萨染色计数肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中巨噬细胞、中性粒细胞、淋巴细胞和嗜酸性粒细胞的数目;ELISA法检测BALF中白细胞介素-13(interleukin-13,IL-13)、IL-6、IL-10、免疫球蛋白E(immunoglobulin E,IgE)、黏蛋白(mucin,Muc)5ac及Muc5b的水平;实时荧光定量PCR检测肺组织中Muc5ac mRNA及Muc5b mRNA的水平。结果:与对照组比较,模型组哮喘大鼠FEV0.3、FVC和PEF降低,气道阻力值升高;BALF中巨噬细胞、中性粒细胞、淋巴细胞、嗜酸性粒细胞数目及IL-13、IL-6、IgE的水平升高,IL-10的水平降低;Muc5ac、Muc5b蛋白及mRNA水平升高;与模型组比较,小青龙汤各剂量组哮喘大鼠FEV0.3、FVC和PEF升高,气道阻力值降低;BALF中巨噬细胞、中性粒细胞、淋巴细胞、嗜酸性粒细胞数目及IL-13、IL-6、IgE的水平降低,IL-10的水平升高;Muc5ac、Muc5b蛋白及mRNA水平降低,且呈剂量依赖性,差异具有统计学意义(P<0.05)。结论:小青龙汤可有效抑制哮喘大鼠的黏液高分泌状态,减轻气道炎症,缓解哮喘症状,其作用机制可能与抑制黏蛋白Muc5ac、Muc5b的分泌有关。

Objective:To observe the influence of Minor Green Dragon Decoction on mucus hypersecretion and airway inflammation in rats with bronchial asthma.Methods:A total of 60 SPF-grade SD rats were randomly divided into the control group,the model group,the ambroxol group,the high-dose Minor Green Dragon Decoction group(14 g·kg^(-1)),the medium-dose Minor Green Dragon Decoction group(7 g·kg^(-1))and the low-dose Minor Green Dragon Decoction group(3.5 g·kg^(-1)),with 10 rats in each group.Except the control group,other groups of rats established bronchial asthma models.After successful modeling,the rats in each group were given drug intervention.The control group and the model group were given the same volume of normal saline by gavage,once a day for 14 consecutive days.The forced expiratory volume(FEV0.3),forced vital capacity(FVC),peak expiratory flow(PEF)and airway resistance(R)were measured with rat lung function tester.The number of macrophages,neutrophils,lym-phocytes and eosinophils in bronchoalveolar lavage fluid(BALF)was counted by using Wright Giemsa staining.The levels of in-terleukin-13(IL-13),IL-6,IL-10,immunoglobulin E(IgE),mucin(Muc)5ac and Mub5b in BALF were detected by ELISA.The levels of Muc5acmRNA and Muc5b protein and mRNA level in lung tissue were detected by real-time fluorescent quantitative PCR.Results:Compared with the control group,FEV0.3,FVC and PEF in the model group decreased,while airway resistance in-creased.The number of macrophages,neutrophils,lymphocytes and eosinophils and the levels of IL-13,IL-6 and IgE in BALF in-creased,but the level of IL-10 decreased;Muc5ac Muc5b protein and mRNA levels increased.Compared with the model group,the FEV0.3,FVC and PEF of asthma rats in each dose group of Minor Green Dragon Decoction increased,while the airway resistance value decreased.The number of macrophages,neutrophils,lymphocytes and eosinophils and the levels of IL-13,IL-6 and IgE in BALF decreased,but the level of IL-10 increased;Muc5ac,Muc5b protein and mRNA levels decreased in a dose-dependent man-ner,and all the differences were statistically significant(P<0.05).Conclusion:Minor Green Dragon Decoction can effectively in-hibit mucus hypersecretion,reduce airway inflammation and relieve asthma symptoms in asthmatic rats.Its effect may be related to the inhibition of mucin Muc5ac and Muc5b secretion.