Cigarette smoke extract inhibits cell migration and contraction via the reactive oxygen species/adenosine monophosphate–activated protein kinase pathway in nasal fibroblasts

• Published in: International forum of allergy & rhinology Vol. 10; no. 3; pp. 356 - 363
• Main Authors: Shin, Jae‐Min, Park, Joo‐Hoo, Yang, Hyun‐Woo, Lee, Heung‐Man, Park, Il‐Ho
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.03.2020
• Subjects: Acetylcysteine; Adenosine; Adenosine kinase; Adult; AMP; AMP-Activated Protein Kinases - metabolism; Cell adhesion & migration; Cell migration; Cell Movement - drug effects; Cell proliferation; Cells, Cultured; Cigarette smoke; cigarette smoke extract; Collagen; Collagen - metabolism; Contractility; Contraction; Female; fibroblast; Fibroblasts; Fibroblasts - drug effects; Fibroblasts - metabolism; Fibroblasts - pathology; Humans; Kinases; Male; Metformin; migration; Nicotiana - chemistry; Phosphorylation; Phosphorylation - drug effects; Protein kinase; Reactive oxygen species; Reactive Oxygen Species - metabolism; Signal transduction; Signal Transduction - drug effects; siRNA; Smoke - adverse effects; Surgery; Turbinates - pathology; Turbinates - surgery; Wound healing; migration; wound healing; cigarette smoke extract; fibroblast; reactive oxygen species
• ISSN: 2042-6976; 2042-6984
• DOI: 10.1002/alr.22479

Background Fibroblast migration plays a significant role in wound healing after endoscopic sinonasal surgery. Cigarette smoke extract (CSE) is a potent inhibitor of fibroblast functions including cell proliferation and migration. The purpose of the study was to determine the influence of CSE on migration and collagen gel contraction in nasal fibroblasts and investigate its underlying mechanisms. Methods Fibroblast migration was evaluated using wound healing assay and transwell migration assay. Contractile activity was assessed by collagen gel contraction assay. Reactive oxygen species (ROS) were quantified by 2′,7′‐dichlorofluorescein diacetate. Fibroblasts were treated with CSE and N‐acetylcysteine (NAC), metformin, compound C, or transfected with small interfering RNA (siRNA) to suppress adenosine monophosphate–activated protein kinase (AMPK) expression. AMPK activation was determined by Western blot. Results CSE and metformin were found to significantly reduce the migration and collagen gel contraction activity of nasal fibroblasts. Conversely, pretreatment with NAC and compound C significantly enhanced the migration and collagen gel contraction activity of fibroblasts. ROS production and AMPK phosphorylation were found to be significantly induced by CSE treatment, whereas the activity was inhibited on treatment with NAC, metformin, compound C, or AMPK siRNA. Silencing of AMPK expression was found to significantly reverse the suppressive effect of CSE in nasal fibroblasts. Conclusion CSE has an inhibitory effect on cell migration and collagen gel contraction activity via the ROS/AMPK signaling pathway in nasal fibroblasts.