中国免疫学杂志2024，Vol.40Issue(10)：2042-2045,2051,5.DOI:10.3969/j.issn.1000-484X.2024.10.004
HNE诱导下PKCδ/θ-Duox1-ROS对气道黏液高分泌影响的体外实验研究
Effect of HNE-induced PKCδ/θ-Duox1-ROS on airway mucus hypersecretion:A vitro experimental study
摘要
Abstract
Objective:To investigate regulatory effect and mechanism of protein kinase C(PKC)δ/θ-dual functional oxidase 1(Duox1)-reactive oxygen species(ROS)signaling pathway on human airway mucin(MUC)5AC,to provide a new target for treatment of high secretion of airway mucus.Methods:Human airway epithelial cells 16HBE were pretreated with PKC and its subunit PKCδ/θ inhibitor,Duox1 inhibitor or free radical scavenger DMTU,respectively,and then human neutrophil elastase(HNE)stimulation to establish an in vitro airway inflammatory cell model.Generation level of ROS in each group of cells was determined by kit,mRNA levels of Duox1 and MUC5AC were detected by real-time fluorescent quantitative PCR,influence of interfering factors of each group of cells on Duox1 protein level was determined by Western blot,and protein expression of MUC5AC in each group of cells was detected by ELISA and immunofluorescence.Results:Compared with control group,ROS production in HNE group was increased significantly,expressions of Duox1 and MUC5AC mRNA and protein were also increased(P<0.05).After administration of Duox1 inhibitors,free radical scavengers or PKC inhibitors and PKCδ/θ inhibitors,ROS production was significantly inhibited,Duox1 and MUC5AC mRNA and protein expressions were decreased(P<0.05),while after giving PKCα/β,ROS generation,Duox1 and MUC5AC mRNA and pro-tein expressions were not significantly changed compared with HNE group(P>0.05).Conclusion:HNE can mediate high expression of MUC5AC through PKCδ/θ-Duox1-ROS,which plays an important role in development of high secretion of airway mucus in vitro cell model experiment.关键词
蛋白激酶Cδ/θ/双重功能氧化酶1/活性氧/黏蛋白5ACKey words
Protein kinase Cδ/θ/Dual oxidase-1/Reactive oxygen species/MUC5AC分类
医药卫生引用本文复制引用
何明欣,杨雅楼,徐立,杨雨菡,张紫薇,周向东,李琪..HNE诱导下PKCδ/θ-Duox1-ROS对气道黏液高分泌影响的体外实验研究[J].中国免疫学杂志,2024,40(10):2042-2045,2051,5.基金项目
国家自然科学基金(82160012,82260001) （82160012,82260001）
国家卫生健康委员会热带病防治重点实验室开放课题(2021NHCTDCKFKT21008) （2021NHCTDCKFKT21008）
海南省重点研发项目(ZDYF2020223) （ZDYF2020223）
海南省重大科技专项(ZDKJ2021036) （ZDKJ2021036）
海南省创新团队项目(820CXTD448) （820CXTD448）
海南省重点研发计划国际科技合作项目(GHYF2022011) （GHYF2022011）
海南省临床医学中心建设项目. （）
