摘要
目的:探讨清肺化痰汤对慢性阻塞性肺疾病(chronic obstructive pulmonary disease, COPD)大鼠模型气道炎症和黏液分泌的调控机制研究。方法:建立COPD大鼠模型。50只大鼠随机分为对照组、COPD模型组和清肺化痰汤低、中、高剂量组。Real-time RT-PCR法检测肺组织MUC5AC mRNA的表达,免疫组织化学法检测肺组织MUC5AC蛋白表达,ELISA检测肺泡灌洗液中MUC5AC的表达,ELISA检测肺组织TNF-α和IL-1β的表达,Western Blot检测肺组织中NF-κB的表达。结果:(1)COPD模型组大鼠肺组织中MUC5AC mRNA明显升高,和正常对照组比较有统计学差异(P<0.05),清肺化痰汤治疗后MUC5AC mRNA的表达水平下降,呈剂量依赖性,和模型组相比较,中剂量组和高剂量组差异有统计学意义;(2)免疫组化结果显示,对照组呈弱阳性表达,COPD模型组可见大量的MUC5AC表达,清肺化痰汤各组治疗后MUC5AC的表达明显下降;(3)ELISA结果显示COPD模型组大鼠肺泡灌洗液中MUC5AC蛋白量明显高于正常对照组(P<0.05);清肺化痰汤各组治疗后,肺泡灌洗液中MUC5AC蛋白量明显低于COPD模型组(P<0.05);(4)ELISA结果显示COPD模型组大鼠肺组织中TNF-α和IL-1β蛋白量明显高于对照组(P<0.05);清肺化痰汤各组治疗后,大鼠肺组织中TNF-α和IL-1β蛋白量明显低于COPD模型组(P<0.05),呈剂量依赖性;(5)Western Blot结果显示COPD模型组大鼠肺组织中NF-κB表达明显高于对照组(P<0.05);清肺化痰汤各组治疗后,大鼠肺组织中NF-κB的表达水平低于COPD模型组(P<0.05),呈剂量依赖性。结论:清肺化痰汤通过抑制NF-κB信号通路调节TNF-α、IL-1β炎症因子的表达,从而抑制气道上皮MUC5AC的表达。
Objective:To investigate the effect of Qingfei Huatan Decoction on mucus hypersecretion in rats with COPD and its mechanism.Methods:The rats model of COPD was established.Fifty rats were randomly divided into control group,model group and Qingfei Huatan Decoction low,medium and high dose groups.Real-time RT-PCR method was used to detect lung tissue MUC5 AC mRNA and the expression of MUC5 AC protein of lung tissue was detected by immune histochemical method.ELISA was used to detect the expression of MUC5 AC in alveolar lavage fluid,the expressions of TNF-αand IL-1βin lung tissue.Western blot detected NF-κB expression in lung tissue.Results:(1)MUC5 AC mRNA was increased obviously in the COPD model group rats lung tissue.Compared with the control group,the difference was significant(P<0.05).Compared with the model group,after Qingfei Huatan Decoction,the expression of MUC5 AC mRNA was descendent.(2)The results of immunohistochemical detection showed that control group’s expression of MUC5 AC was weakly positive.COPD model group had a lot o expression of MUC5 AC.The expression of MUC5 AC was declined obviously after Qingfei Huatan Decoction treatment.(3)ELISA showed that MUC5 AC protein of alveolar lavage in the rats of the COPD model group was significantly higher than that of the normal control group(P<0.05).Qingfei Huatan Decoction group’s alveolar lavage fluid MUC5 AC protein was significantly lower than that of the COPD model group(P<0.05).(4)ELISA showed that the proteins of TNF-αand IL-1βin the COPD model group rats lung tissue were significantly higher than those of the control group(P<0.05).Qingfei Huatan Decoction group’s proteins of TNF-αand IL-1βin rats lung tissue were significantly lower than those of the COPD model group(P<0.05).(5)Western blot results showed that the expression of NF-κB in COPD model group rat lung tissue was significantly higher than that of the control group(P<0.05).Qingfei Huatan Decoction group’s expression of the NF-κB level in lung tissue of rats was lower than that of the COPD model group(P<0.05).Conclusion:Qingfei Huatan Decoction can inhibit the expression of MUC5 AC,and its mechanism may be related to Qingfei Huatan Decoction depressing the expressions of TNF-α,IL-1βand NF-κB.
作者
屈飞
黄小婷
崔艳茹
李惠兰
邱沙英
QU Fei;HUANG Xiaoting;CUI Yanru;LI Huilan;QIU Shaying(Dept of Pharmacology,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,Jiangxi,China;Key Laboratory of pharmacology of Traditional Chinese Medicine in Jiangxi,Nanchang 330004,Jiangxi,China;Dept of Physiology,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004,Jiangxi,China)
出处
《中华中医药学刊》
CAS
北大核心
2019年第9期2064-2067,2305,共5页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金项目(81760744)
江西省自然科学基金项目(20151BAB215036)
江西省卫生厅中医药科研基金项目(2012A154,2015A038)
江西省教育厅科技项目(150870)
作者简介
屈飞(1980-),男,江西九江人,副教授,硕士研究生导师,博士,研究方向:中药药理学研究;通讯作者:崔艳茹(1980-),女,河北保定人,副教授,硕士研究生导师,硕士,研究方向:中药药理学研究。
