基于EGFR/MAPK/MUC5AC通路探讨益肺健脾方对慢性阻塞性肺疾病模型大鼠气道黏液高分泌的影响  

To Explore the Effect of Yifei Jianpi Prescription on Airway Mucus Hypersecretion in COPD Model Rats Based on EGFR/MAPK/MUC5AC Pathway

作　　者：张雅男 张玉新 郑莉莉 王胜[3] ZHANG Yanan;ZHANG Yuxin;ZHENG Lili;WANG Sheng(The First Clinical Medical College of Anhui University of Chinese Medicine,Hefei 230012 Anhui,China;Lu’an Hospital of Traditional Chinese Medicine,Lu’an 237000 Anhui,China;Department of Respiratory Medicine,Geriatric Center,the First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031 Anhui,China)

机构地区：[1]安徽中医药大学第一临床医学院,安徽合肥230012 [2]六安市中医院,安徽六安237000 [3]安徽中医药大学第一附属医院老年病中心呼吸内科,安徽合肥230031

出　　处：《中药新药与临床药理》2024年第8期1115-1122,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(82274500);安徽高校自然科学研究重点项目(KJ2020A0425);安徽省高等学校自然科学研究重点项目(2022AH050493);王胜安徽省名中医工作室项目。

摘　　要：目的基于表皮生长因子受体/丝裂原活化蛋白激酶/黏蛋白5AC(EGFR/MAPK/MUC5AC)通路探析益肺健脾方对慢性阻塞性肺疾病(COPD)大鼠气道黏液高分泌的治疗作用及机制。方法将SD雄性大鼠随机分为空白组、模型组、氨溴索组及益肺健脾方低、中、高剂量组。烟熏联合脂多糖滴注、番泻叶灌胃泻下的方式复制肺脾两虚型COPD大鼠模型。阿利新蓝-过碘酸-雪夫染色(AB-PAS)观察大鼠支气管黏液分泌及杯状细胞增生程度;苏木精-伊红染色(HE)观察大鼠肺组织病理形态;蛋白质免疫印迹法(Western Blot)检测大鼠肺组织MUC5AC、EGFR蛋白和c-Jun氨基末端激酶(JNK)、p38丝裂活化蛋白激酶(p38 MAPK)、细胞外调节蛋白激酶(ERK)蛋白及磷酸化蛋白的表达;采用实时荧光定量PCR法(RT-qPCR)检测大鼠肺组织EGFR、JNK、p38 MAPK、MUC5AC及ERK mRNA的表达。结果AB-PAS染色显示氨溴索组及益肺健脾方各剂量组杯状细胞增生明显改善。HE染色下见氨溴索组及益肺健脾方高剂量组明显改善肺组织病理损伤。相较于空白组,模型组肺组织中ERK、MUC5AC、p38 MAPK、JNK及EGFR mRNA的表达皆明显升高(P<0.01),MUC5AC、EGFR蛋白和p38 MAPK、JNK、ERK蛋白及磷酸化蛋白表达量皆明显升高(P<0.01)。相比于模型组,氨溴索组及益肺健脾方中、高剂量组各蛋白表达量明显下降(P<0.01,P<0.05),低剂量组EGFR、p38 MAPK、p-p38 MAPK蛋白表达量皆降低(P<0.01,P<0.05);高剂量组与氨溴索组EGFR、ERK、JNK、p38 MAPK及MUC5AC mRNA的表达皆降低(P<0.01,P<0.05),中、低剂量组MUC5AC mRNA的表达下降(P<0.05)。结论益肺健脾方可减少MUC5AC的表达,从而改善气道黏液高分泌,其作用机制与EGFR/MAPK信号通路有关。Objective To explore the therapeutic effects and mechanism of Yifei Jianpi Prescription on airway mucus hypersecretion in chronic obstructive pulmonary disease(COPD)rats through the epidermal growth factor receptor/mitogen-activated protein kinase/mucin 5AC(EGFR/MAPK/MUC5AC)pathway.Methods Male SD rats were randomly divided into blank group,model group,ambroxol group and Yifei Jianpi Prescription low-,medium-and high-dose groups.The COPD rat model with lung and spleen deficiency was replicated by smoking combined with lipopolysaccharide instillation and administration of Folium Sennae.Alcian blue-periodic acid-Schiff staining(AB-PAS)was used to observe bronchial mucus secretion and the degree of goblet cell hyperplasia in rats.Hematoxylin-eosin staining(HE)was used to observe the pathological morphology of lung tissue of rats.Western Blot was used to detect the expression of MUC5AC,EGFR protein,c-Jun N-terminal kinase(JNK),p38 MAPK,protein and phosphorylated protein of extracellular regulated protein kinase(ERK)in rat lung tissues.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the mRNA expression of EGFR,JNK,p38 MAPK,MUC5AC,and ERK in rat lung tissues.Results AB-PAS staining showed that the number of goblet cells decreased significantly in ambroxol group and all dose groups of Yifei Jianpi Prescription.HE staining showed that ambroxol group and Yifei Jianpi high-dose group significantly improved the pathological injury of lung tissue.Compared with the blank group,the mRNA expressions of ERK,MUC5AC,p38 MAPK,JNK and EGFR were significantly increased in the lung tissues of the model group(P<0.01),and the expression of MUC5AC and EGFR protein,as well as p38 MAPK,JNK,ERK total and phosphorylated protein were significantly increased(P<0.01).Compared with model group,the protein expression levels of ambroxol group and Yifei Jianpi Prescription medium-and high dose groups were significantly decreased(P<0.01,P<0.05),while the protein expression levels of EGFR,p38 MAPK and p-p38 MAPK in Yifei Jianpi Pre

关 键 词：慢性阻塞性肺疾病 肺脾两虚 MUC5AC EGFR MAPK 益肺健脾方 气道黏液 大鼠 

分 类 号：R285.5[医药卫生—中药学]