title: Notch signaling regulates col1α1 and col1α2 expression in airway fibroblasts.
abstract: Subepithelial fibrosis is one of the common pathological features of asthmatic airway remodeling. During subepithelial fibrosis, type I collagen becomes the most abundant extracellular protein component. Studies have shown that Notch signaling participates in the progression of fibrosis; however, whether Notch signaling is involved in regulating type I collagen expression in airway fibroblasts remains unclear. The aim of the present study was to examine whether Notch signaling can regulate type I collagen expression in airway fibroblasts and to explore the underlying molecular mechanisms. Here, the expression of Notch signaling components was examined in mouse L929 cells and human MRC-5 cells. After upregulating or downregulating Notch signaling in these cell lines, col1α1 and col1α2 expression was examined. Using gene reporter assays, site-directed mutagenesis, and ChIP assays, the role of Hes1 binding sites in both the mouse and human COL1A1 and COL1A2 promoters was investigated. This study revealed that Notch signaling-related molecules (including Notch1, Hes1, and others) are expressed in L929 and MRC-5 cells and that Notch signaling regulates the expression of col1α1 and col1α2 in both cell lines. Additionally, over-expression of the Notch intracellular domain resulted in activation of the COL1A1 and COL1A2 promoters, and site-directed mutagenesis reporter assays revealed that Hes1 proteins might augment both mouse and human COL1A1 and COL1A2 promoter activity. Furthermore, ChIP assays confirmed that Hes1 binds to the COL1A1 and COL1A2 promoters in both L929 and MRC-5 cells. Therefore, it is reasonable to assume that Notch signaling can directly upregulate COL1A1 and COL1A2 promoter activity through a Hes1-dependent mechanism, which could serve as a possible target for pharmacotherapy of airway subepithelial fibrosis.
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xincheng District, Xi'an 710032, China Department of Respiratory Medicine, 306th Hospital of PLA, Chaoyang District, Beijing 100101, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Hu
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xincheng District, Xi'an 710032, China.
authors.forename: Hai-Feng
authors.initials: HF
authors.lastname: Ou-Yang
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xincheng District, Xi'an 710032, China changguinew@126.com.
authors.forename: Chang-Gui
authors.initials: CG
authors.lastname: Wu
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xincheng District, Xi'an 710032, China.
authors.forename: Shuo-Yao
authors.initials: SY
authors.lastname: Qu
authors.affiliation: Department of Respiratory Medicine, Xijing Hospital, Fourth Military Medical University, Xincheng District, Xi'an 710032, China.
authors.forename: Xin-Ting
authors.initials: XT
authors.lastname: Xu
authors.affiliation: Department of Respiratory Medicine, 306th Hospital of PLA, Chaoyang District, Beijing 100101, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Wang
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D001665:Binding Sites; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D024042:Collagen Type I / Q000096:biosynthesis*; D005347:Fibroblasts / Q000502:physiology*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051880:Receptors, Notch / Q000378:metabolism*; D015398:Signal Transduction; D000072056:Transcription Factor HES-1
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D024042:Collagen Type I; D018398:Homeodomain Proteins; D051880:Receptors, Notch; D000072056:Transcription Factor HES-1; C489734:Hes1 protein, mouse; C078493:HES1 protein, human
keywords: Notch; asthma; col1α1; col1α2; subepithelial fibrosis