目的:研究复方银翘合剂对哮喘小鼠的作用,并探讨其可能的作用机制.方法:将72 只小鼠随机分为6 组,正常组、模型组、复方银翘合剂低、中、高剂量组和阳性药物组,每组12 只.采用卵清蛋白诱导小鼠哮喘模型,观察小鼠一般症状并检测IgE浓度.收集小鼠肺泡灌洗液并染色计数,HE染色观察小鼠肺组织病理变化.ELISA检测小鼠肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-4、IL-5、IL-13 的水平.Western blot 法检测肺组织中 Toll 样受体 4(TLR4)、髓样分化因子 88(MyD88)和核转录因子κB(NF-κB)蛋白表达水平.结果:与正常组比较,模型组小鼠肺泡灌洗液中总细胞数、嗜酸性粒细胞、淋巴细胞、中性粒细胞均显著增多(P<0.05),巨噬细胞显著减少(P<0.05),肺组织中支气管管壁增厚,炎性细胞浸润,TNF-α、IL-4、IL-5、IL-13 的水平均显著提高(P<0.05),TLR4、MyD88 和NF-κB蛋白表达水平均显著升高(P<0.05).与模型组比较,复方银翘合剂中、高剂量组小鼠肺泡灌洗液中总细胞数、嗜酸性粒细胞、淋巴细胞、中性粒细胞均显著减少(P<0.05),巨噬细胞显著增多(P<0.05),TNF-α、IL-4、IL-5、IL-13 的水平均显著降低(P<0.05),TLR4、MyD88 和NF-κB蛋白表达水平均显著降低(P<0.05).复方银翘合剂各剂量组比较结果显示,高剂量组的上述变化最明显.结论:复方银翘合剂可缓解哮喘小鼠症状,减轻肺组织病理变化和炎症水平,其作用机制可能与抑制TLR4/MyD88/NF-κB信号通路有关.
Objective:To study the effect of compound lonicera and forsythia mixture(CLFM)on asthmatic mice and its possible mechanism.Methods:72 mice were divided into 6 groups:normal group,model group,positive drug group,and low,medium and high dose of CLFM groups,with 12 mice in each group.The asthma model was induced by ovalbumin,the general symptoms of mice were observed,and IgE concentration was measured.The bronchoalveolar lavage fluid of mice was collected and counted by staining.The pathological changes of lung tissue were observed by HE staining.The levels of tumor necrosis factor(TNF)-α,interleukin(IL)-4,IL-5 and IL-13 in bronchoalveolar lavage fluid were determined by ELISA.The protein expression levels of toll-like receptor-4(TLR4),myeloid differentiation factor 88(MyD88)and nuclear factor kappa B(NF-κB)were detected by Western blot.Results:Compared with the normal group,the total number of cells,eosinophils,lymphocytes and neutrophils in the bronchoalveolar lavage fluid of the model group were significantly increased(P<0.05),the number of macrophages was significantly decreased(P<0.05),the bronchial wall in the lung tissue was thickened with inflammatory cell infiltration.The levels of TNF-α,IL-4,IL-5 and IL-13 were significantly increased(P<0.05),and the protein expression levels of TLR4,MyD88 and NF-κB were significantly increased(P<0.05).Compared with the model group,the total number of cells,eosinophils,lymphocytes and neutrophils in bronchoalveolar lavage fluid in the medium and high dose of CLFM groups were significantly decreased(P<0.05),and the number of macrophages was significantly increased(P<0.05).The levels of TNF-α,IL-4,IL-5 and IL-13 were significantly decreased(P<0.05),and the protein expression levels of TLR4,MyD88 and NF-κB were significantly increased(P<0.05).The comparison results of different dose groups of CLFM showed that the above changes were most pronounced in the high-dose group.Conclusion:Compound lonicera and forsythia mixture can alleviate the symptoms of asthmatic mice,alleviate the pathological changes and inflammatory level of lung tissue,and its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway.