摘要
目的探讨烟曲霉提取物(AFE)对人支气管上皮细胞(16HBE)黏蛋白5AC(MUC5AC)表达的影响及其可能的信号转导通路。方法本研究为实验研究。将16HBE分为4组:正常溶剂对照组、AFE处理组、AFE+选择性EGFR酪氨酸激酶抑制剂(AG1478)作用组和AFE+MAPK激酶抑制剂(PD98059)作用组。用免疫荧光、免疫组织化学、RT-PCR、Western blot及ELISA等方法测定EGFR、ERK1/2、磷酸化EGFR、磷酸化ERK1/2及MUC5AC的表达。结果8~24 mg/L AFE与16HBE共孵育24 h,各组细胞EGFR相对灰度值分别为0.4±0.1、1.0±0.4、1.3±0.3、1.8±0.8,ERK1/2相对灰度值分别为1.2±0.4、2.4±0.5、3.2±0.7、4.3±0.8,可剂量依赖性上调EGFR和ERK1/2表达;16 mg/L AFE与16HBE共孵育1 h,磷酸化EGFR的相对灰度值为1.2±0.2,磷酸化ERK1/2的相对灰度值为2.23±0.50,可诱导EGFR和ERK1/2磷酸化;16HBE与10μmol/L的AG1478作用可抑制AFE诱导的EGFR和ERK1/2的磷酸化,与30μmol/L的MAPK激酶抑制剂PD98059作用可抑制ERK1/2的磷酸化,而不影响EGFR的磷酸化。16 mg/L的AFE可上调16HBE MUC5AC的表达,MUC5AC mRNA光密度比值为2.2±0.2,与AG1478或PD98059作用可抑制AFE诱导MUC5AC表达。结论烟曲霉可上调16HBE细胞MUC5AC的表达,激活EGFR-ERK1/2信号通路是烟曲霉上调16HBE细胞MUC5AC的表达机制之一。
Objective To explore the effects and possible signal pathways of Aspergillus fumigatus extract(AFE)on the expression of Mucin 5A and C(MUC5AC)in human bronchial epithelial cells.Methods This was an experimental study.The 16HBE cells were divided into 4 groups:normal medium control group,AFE-treated group,AFE+selective inhibitor of protein tyrosine kinase of EGFR(AG1478)treated group,and AFE+inhibitor of MAPK kinase(PD98059)treated group.The expression of EGFR,extracellular-signal regulated kinase 1/2(ERK1/2),phospho-EGFR,phospho-ERK1/2,and MUC5AC were measured by immunofluorescence,immunohistochemistry,RT-PCR,Western blot,and ELISA.Results Incubation with 16HBE cells for 24 h,8-24 mg/L of AFE dose-dependently upregulated the expression of EGFR and ERK1/2.The relative gray values of EGFR were 0.4±0.1,1.0±0.4,1.3±0.3,and 1.8±0.8,and the relative gray values of ERK1/2 were 1.2±0.4,2.4±0.5,3.2±0.7,and 4.3±0.8,respectively.Incubation with 16HBE cells for 1 h,16 mg/L of AFE induced phosphorylation of EGFR and ERK1/2.The relative gray value of phospho-EGFR was 1.2±0.2,and that of phospho-ERK1/2 was 2.23±0.50.Incubation with 16HBE cells,10μmol/L of AG1478 inhibited the phosphorylation of EGFR and ERK1/2,whereas 30μmol/L of PD98059 only inhibited the phosphorylation of ERK1/2 and had no influence on the phosphorylation of EGFR.16 mg/L of AFE upregulated the expression of MUC5AC in 16HBE cells,The optical density ratio of MUC5AC mRNA was 2.2±0.2.The expression of MUC5AC induced by AFE in 16HBE cells could be inhibited after incubation with AG1478 or PD98059.Conclusions Aspergillus fumigatus can increase the expression of MUC5AC in 16HBE cells.Activation of EGFR-ERK1/2 signal pathway is one of the mechanisms by which Aspergillus fumigatus upregulates the expression of MUC5AC in 16HBE cells.
作者
王妍竹
李萌
高福生
Wang Yanzhu;Li Meng;Gao Fusheng(Affiliated Hospital of Weifang Medical University,School of Clinical Medicine,Weifang Medical University,Weifang 261000,China;School of Clinical Medicine,Weifang Medical University,Weifang 261000,China;Department of Respiratory Medicine,the Affiliated Hospital of Weifang Medical College,Weifang 261000,China)
出处
《国际呼吸杂志》
2023年第6期679-686,共8页
International Journal of Respiration
关键词
哮喘
气道重塑
烟曲霉
黏蛋白5AC
表皮生长因子受体
Asthma
Airway remodeling
Aspergillus fumigatus
Mucin 5A and C
Epidermal growth factor receptor