摘要
目的:观察固本防哮饮对哮喘缓解期小鼠模型Toll-样受体(TLR) 4,TLR7,核转录因子-κB p65(NF-κB p65),NF-κB抑制蛋白α(IκBα)的调节作用,探讨固本防哮饮治疗哮喘缓解期的机制。方法:呼吸道合胞病毒(RSV)联合鸡卵蛋白(OVA)对3周龄BALB/c小鼠制备哮喘缓解期模型,将60只小鼠分为空白组、模型组、地塞米松组(0. 001 g·kg^(-1))以及固本防哮饮低、中、高剂量(6. 5,13,26 g·kg^(-1))组,分别进行干预,共给药28 d,每天1次。给药结束后处死小鼠,苏木素-伊红(HE)染色观察哮喘小鼠肺组织病理形态学变化,并对肺部炎症情况进行评分;蛋白免疫印迹法(Western blot)检测小鼠肺组织TLR4,TLR7,IκBα及肺组织细胞核内NF-κB p65蛋白表达水平;免疫荧光共聚焦观察NF-κB p65在细胞核表达情况。结果:与空白组比较,模型组小鼠肺组织有明显炎症细胞浸润及支气管狭窄(P <0. 01),同时小鼠肺组织细胞核内NF-κB p65分布显著增多(P <0. 01),表达明显增高(P <0. 05),TLR4,TLR7,IκBα蛋白表达明显降低(P <0. 05);与模型组比较,固本防哮饮各剂量组能显著缓解小鼠肺组织炎症浸润(P <0. 01),降低肺组织核蛋白NF-κB p65的表达(P <0. 01),核蛋白NF-κB p65入核情况减轻(P <0. 01),同时显著增加TLR4,TLR7,IκBα蛋白的表达(P <0. 01)。结论:固本防哮饮能够缓解气道炎症,并且可能是通过调节TLR4,TLR7,NF-κB p65,IκBα实现其治疗作用。
Objective:To observe the regulatory effect of Guben Fangxiao decoction on Toll-like receptor(TLR)4,TLR7,nuclear factor-kappa B p65(NF-κB p65)and NF-κB inhibitor protein alpha(IκBα)in mice with asthma remission,in order to explore the mechanism of Guben Fangxiao decoction in treating asthma remission.Method:Respiratory syncytial virus(RSV)combined with chicken ovalbumin(OVA)was used to build asthma remission model in 3-week-old BALB/c mice.Sixty mice were divided into blank group,model group,dexamethasone group(0.001 g·kg-1),low,medium and high-dose Guben Fangxiao decoction group(6.5,13,26 g·kg-1),respectively.Intervention was given once a day for 28 days.After administration,the mice were put to death.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of lung tissue and score the pulmonary inflammation.Western blot was used to detect the expressions of TLR4,TLR7,IκBαand nuclear NF-κB p65 in lung tissue of mice.Immunofluorescence was used to observe the expression of NF-κB p65 in cellnucleuses.Result:Compared with blank group,the lung tissue of model group showed obvious inflammatory cell infiltration(P<0.01)and bronchial stenosis,the nuclear distribution of NF-κB p65 increased significantly(P<0.01),and the expressions of TLR4,TLR7 and IκBαproteins increased significantly(P<0.05).Compared with the model group,Guben Fangxiao decoction could significantly alleviate the inflammatory infiltration of lung tissue in mice(P<0.01),reduce the expression of nuclear protein NF-κB p65(P<0.01),relieve the nuclear entry of nuclear protein NF-κB p65(P<0.01),and increase the expressions of TLR4,TLR7 and IκBαproteins(P<0.01).Conclusion:Guben Fangxiao decoction can alleviate airway inflammation,and its therapeutic effect may be achieved by regulating TLR4,TLR7,NF-κB p65 and IκBα.
作者
卓月
袁雪晶
纪建建
ZHUO Yue;YUAN Xue-jing;JI Jian-jian(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第10期57-63,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
江苏省"六大人才高峰"高层次人才B类资助项目(2016-WSN-059)