TGF-\u03b21-induced HSP47 regulates extracellular matrix accumulation via Smad2/3 signaling pathways in nasal fibroblasts

Hae-Ji Kim,Jae-Min Shin,Il-Ho Park,Heung-Man Lee,Hyun-Woo Yang,Joo-Hoo Park

doi:10.1038/s41598-019-52064-1

Hae-Ji Kim, Jae-Min Shin + Show 4 more

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https://doi.org/10.1038/s41598-019-52064-1

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Abstract

HSP47 is required for the production of collagen and serves an important role in tissue remodeling, a pathophysiologic mechanism of chronic rhinosinusitis (CRS). We investigated the relationship between HSP47 expression and tissue remodeling in CRS. We also determined the underlying molecular mechanisms of TGF-β1-induced HSP47 and extracellular matrix (ECM) production in nasal fibroblasts. HSP47, α-SMA, fibronectin, and collagen type I expression levels were measured using real-time PCR, western blotting, and immunofluorescence staining. Fibroblast migration was analyzed using scratch and transwell migration assays. Contractile activity was measured with a collagen gel contraction assay. HSP47 is increased in patients with CRS without nasal polyps. TGF-β1 induced HSP47 expression in nasal fibroblasts. Myofibroblast differentiation and ECM production, which are induced by TGF-β1, were inhibited by siHSP47. We also confirmed that the Smad2/3 signaling pathway is involved in TGF-β1-induced HSP47 expression in nasal fibroblasts. In a functional assay, TGF-β1-enhanced migration and contraction ability were inhibited by HSP47 knockout. Glucocorticoid reversed the stimulatory effects of TGF-β1 on HSP47 expression and ECM production in nasal fibroblasts and ex vivo organ cultures. HSP47 expression is involved in TGF-β1-induced myofibroblast differentiation and ECM production through the Smad2/3 signaling pathway, which might contribute to tissue remodeling in chronic rhinosinusitis.

Highlights

Heat shock protein 47 (HSP47) is required for the production of collagen and serves an important role in tissue remodeling, a pathophysiologic mechanism of chronic rhinosinusitis (CRS)
We analyzed whether it was possible to distinguish between patients with chronic rhinosinusitis with nasal polyp formation (CRSwNP) or chronic rhinosinusitis without nasal polyps (CRSsNP) based on severity using the Rhinosinusitis Lund-McKay CT scores
Protein levels of HSP47 and extracellular matrix (ECM)-related markers were determined by western blotting. (C) Nasal fibroblasts transfected with HSP47 siRNA were stimulated with or without TGF-β1 for 48 h

Summary

Introduction

HSP47 is required for the production of collagen and serves an important role in tissue remodeling, a pathophysiologic mechanism of chronic rhinosinusitis (CRS). HSP47 expression is involved in TGF-β1-induced myofibroblast differentiation and ECM production through the Smad2/3 signaling pathway, which might contribute to tissue remodeling in chronic rhinosinusitis. A dynamic process involving the production and degradation of the extracellular matrix (ECM), is important in the process of wound healing for all organs of the human body, and this sometimes involves pathological tissue regeneration[5] Chronic airway diseases such as asthma and CRS stimulate mechanisms of airway healing and repair that can lead to progressive, potentially irreversible, tissue destruction. Fibroblasts play a role in structurally maintaining tissue morphology and inducing ECM production During this process, fibroblasts develop into myofibroblasts that express α-smooth muscle actin (α-SMA), subsequently resulting in increased deposition of ECM components, such as fibronectins and collagen type I7. Myofibroblasts, types of HSP47-positive cells, are proposed to play a key role in the synthesis, deposition, and remodeling of the ECM in nasal fibrosis in human patients and animal models[11]

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