黄龙止咳口服液对咳嗽变异性哮喘模型小鼠的作用机制研究

Study on the mechanism of Huanglong Zhike Oral Liquid on mouse with cough variant asthma model

关键词:  黄龙止咳口服液  咳嗽变异性哮喘  气道炎症  Toll样受体  核转录因子-κB

Key words:  Huanglong Zhike Oral Liquid  cough variant asthma  airway inflammation  Toll-like receptors  NF-κB

摘要:

[目的] 探讨黄龙止咳口服液治疗咳嗽变异性哮喘的可能作用机制。[方法] 将50只BALB/c小鼠随机分为正常组、模型组、地塞米松组、黄龙止咳口服液低、高剂量组，每组10只。采用卵蛋白、氢氧化铝腹腔注射致敏并吸入卵蛋白激发法建立咳嗽变异性哮喘小鼠模型，黄龙止咳口服液组分别以0.75 g/kg和1.5 g/kg进行灌胃，每日1次，持续14 d后处死小鼠。采集血清，以酶联免疫吸附（ELISA）法检测血清总免疫球蛋白E（IgE）及白细胞介素（IL）-4、IL-5、IL-13的含量；分离小鼠肺组织，苏木精-伊红（HE）染色后观察肺组织形态；蛋白免疫印迹（Western blot）法检测肺组织中Toll样受体4（TLR4）、髓样分化因子（MyD88）及核转录因子-κB（NF-κB）的蛋白表达量。[结果] 与正常组比较，模型组血清中各炎症细胞及IgE、IL-4、IL-5、IL-13含量均明显增高，肺组织中TLR4、MyD88和NF-κB蛋白表达增高，差异有统计学意义（P<0.01）。与模型组比较，地塞米松组、黄龙止咳口服液低剂量组和高剂量组病理切片表现均有所改善，血清中各炎症细胞及炎症因子含量、肺组织中TLR4、MyD88和NF-κB蛋白表达均降低（P<0.01）。[结论] 黄龙止咳口服液能改善咳嗽变异性哮喘模型小鼠气道炎症及气道高反应性，其机制可能与抑制TLR4/MyD88/NF-κB信号通路有关。

Abstract:

[Objective] To investigate the possible mechanism of Huanglong Zhike Oral Liquid in the treatment of cough variant asthma.[Methods] Fifty BALB/c mice were randomly divided into control group,the model group,dexamethasone (Dex) group and Huanglong Zhike Oral Liquid low and high dose groups,with 10 mice in each group. The mice model of cough variant asthma was established by intraperitoneal injection of ovalbumin and aluminum hydroxide,sensitization and inhalation of ovalbumin. The mice in Huanglong Zhike Oral Liquid group were given 0.75g/kg and 1.5g/kg by gavage respectively once a day. The mice were killed after 14 days.Serum was collected and the levels of total IgE and interleukin-4 (IL-4),IL-5 and IL-13 were detected by ELISA. Lung tissues of mice were extracted and observed by HE staining. Toll-like receptors 4 (TLR4),myeloid differentiation factor 88 (MyD88)and nuclear factor-κB (NF-κB) were detected by Western blot.[Results] Compared with the control group,the levels of inflammatory cells,IgE,IL-4,IL-5 and IL-13 in the serum of the model group were significantly increased,and the expressions of TLR4,MyD88 and NF-κB in the lung tissue were significantly increased (P<0.01). Compared with the model group,the pathological sections of the Dex group,Huanglong Zhike Oral Liquid low-dose group and high-dose group were improved,the content of inflammatory cells and inflammatory factors in serum,the expression of TLR4,MyD88 and NF-κ B protein in lung tissue were decreased (P<0.01).[Conclusion] Huanglong Zhike Oral Liquid can improve airway inflammation and airway hyperresponsiveness in cough variant asthma model mice,and its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signal pathway.