Role of reactive oxygen species in transforming growth factor beta1-induced alpha smooth-muscle actin and collagen production in nasal polyp-derived fibroblasts

Il Ho Park, Se Jin Park, Jung Sun Cho, You Mi Moon, Tae Hoon Kim, Sang Hag Lee, Heung Man Lee

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Background: Myofibroblasts are detected in nasal polyps and are involved in nasal polyp formation by inducing extracellular matrix accumulation. Reactive oxygen species (ROS) are released during the differentiation of fibroblasts to myofibroblasts. The purpose of this study was to investigate ROS production and nicotinamide adenine dinucleotide phosphate oxidase (NOX) expression in nasal polyp-derived fibroblasts (NPDFs) and to evaluate whether ROS from NOX mediates transforming growth factor (TGF)-β1-induced production of alpha smooth-muscle actin (α-SMA) and collagen production. Methods: NPDFs were incubated and treated with TGF-β1. The mRNA expression of NOXs, α-SMA, and collagen type I and IV was determined by reverse transcription-polymerase chain reaction, and the expression of α-SMA protein was determined by immunofluorescence microscopy. The amount of total soluble collagen production was analyzed by the SirCol assay. The ROS generation of cells was investigated using the 2′,7′-dichlorfluorescein- diacetate. The fluorescence was captured by fluorescent microscope and measured using a fluorometer. Results: Stimulation with TGF-β1 increased ROS production by NPDFs compared with NPDFs not treated with TGF-β1. Stimulation with TGF-β1 increased the expression of NOX4 mRNA most potently among various Nox enzymes. siNOX4 was able to decrease the level of ROS production. Myofibroblast differentiation and the production of collagen in NPDFs were prevented by inhibition of ROS generation with diphenyliodonium, N-acetylcysteine, ebselen, and siNox4. Conclusions: This study showed that NOX4 and ROS have a role in myofibroblast differentiation and collagen production of TGF-β1-induced NPDFs and that these processes are inhibited by the elimination of ROS.

Original languageEnglish
Pages (from-to)278-286
Number of pages9
JournalInternational Archives of Allergy and Immunology
Volume159
Issue number3
DOIs
Publication statusPublished - 2012 Oct 1

Fingerprint

Nasal Polyps
Transforming Growth Factor beta1
Smooth Muscle
Actins
Reactive Oxygen Species
Collagen
Fibroblasts
Transforming Growth Factors
Myofibroblasts
Messenger RNA
Collagen Type IV
Acetylcysteine
Collagen Type I
NADP
Fluorescence Microscopy
Reverse Transcription
Extracellular Matrix
Oxidoreductases
Fluorescence
Polymerase Chain Reaction

Keywords

  • Alpha-smooth-muscle actin
  • Myofibroblast
  • Nasal polyps
  • Nicotinamide adenine dinucleotide phosphate oxidase
  • Reactive oxygen species
  • Transforming growth factor-beta1

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this

@article{fb6d340d99084369b81012ceaae00d78,
title = "Role of reactive oxygen species in transforming growth factor beta1-induced alpha smooth-muscle actin and collagen production in nasal polyp-derived fibroblasts",
abstract = "Background: Myofibroblasts are detected in nasal polyps and are involved in nasal polyp formation by inducing extracellular matrix accumulation. Reactive oxygen species (ROS) are released during the differentiation of fibroblasts to myofibroblasts. The purpose of this study was to investigate ROS production and nicotinamide adenine dinucleotide phosphate oxidase (NOX) expression in nasal polyp-derived fibroblasts (NPDFs) and to evaluate whether ROS from NOX mediates transforming growth factor (TGF)-β1-induced production of alpha smooth-muscle actin (α-SMA) and collagen production. Methods: NPDFs were incubated and treated with TGF-β1. The mRNA expression of NOXs, α-SMA, and collagen type I and IV was determined by reverse transcription-polymerase chain reaction, and the expression of α-SMA protein was determined by immunofluorescence microscopy. The amount of total soluble collagen production was analyzed by the SirCol assay. The ROS generation of cells was investigated using the 2′,7′-dichlorfluorescein- diacetate. The fluorescence was captured by fluorescent microscope and measured using a fluorometer. Results: Stimulation with TGF-β1 increased ROS production by NPDFs compared with NPDFs not treated with TGF-β1. Stimulation with TGF-β1 increased the expression of NOX4 mRNA most potently among various Nox enzymes. siNOX4 was able to decrease the level of ROS production. Myofibroblast differentiation and the production of collagen in NPDFs were prevented by inhibition of ROS generation with diphenyliodonium, N-acetylcysteine, ebselen, and siNox4. Conclusions: This study showed that NOX4 and ROS have a role in myofibroblast differentiation and collagen production of TGF-β1-induced NPDFs and that these processes are inhibited by the elimination of ROS.",
keywords = "Alpha-smooth-muscle actin, Myofibroblast, Nasal polyps, Nicotinamide adenine dinucleotide phosphate oxidase, Reactive oxygen species, Transforming growth factor-beta1",
author = "Park, {Il Ho} and Park, {Se Jin} and Cho, {Jung Sun} and Moon, {You Mi} and Kim, {Tae Hoon} and Lee, {Sang Hag} and Lee, {Heung Man}",
year = "2012",
month = "10",
day = "1",
doi = "10.1159/000337460",
language = "English",
volume = "159",
pages = "278--286",
journal = "International Archives of Allergy and Immunology",
issn = "1018-2438",
publisher = "S. Karger AG",
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TY - JOUR

T1 - Role of reactive oxygen species in transforming growth factor beta1-induced alpha smooth-muscle actin and collagen production in nasal polyp-derived fibroblasts

AU - Park, Il Ho

AU - Park, Se Jin

AU - Cho, Jung Sun

AU - Moon, You Mi

AU - Kim, Tae Hoon

AU - Lee, Sang Hag

AU - Lee, Heung Man

PY - 2012/10/1

Y1 - 2012/10/1

N2 - Background: Myofibroblasts are detected in nasal polyps and are involved in nasal polyp formation by inducing extracellular matrix accumulation. Reactive oxygen species (ROS) are released during the differentiation of fibroblasts to myofibroblasts. The purpose of this study was to investigate ROS production and nicotinamide adenine dinucleotide phosphate oxidase (NOX) expression in nasal polyp-derived fibroblasts (NPDFs) and to evaluate whether ROS from NOX mediates transforming growth factor (TGF)-β1-induced production of alpha smooth-muscle actin (α-SMA) and collagen production. Methods: NPDFs were incubated and treated with TGF-β1. The mRNA expression of NOXs, α-SMA, and collagen type I and IV was determined by reverse transcription-polymerase chain reaction, and the expression of α-SMA protein was determined by immunofluorescence microscopy. The amount of total soluble collagen production was analyzed by the SirCol assay. The ROS generation of cells was investigated using the 2′,7′-dichlorfluorescein- diacetate. The fluorescence was captured by fluorescent microscope and measured using a fluorometer. Results: Stimulation with TGF-β1 increased ROS production by NPDFs compared with NPDFs not treated with TGF-β1. Stimulation with TGF-β1 increased the expression of NOX4 mRNA most potently among various Nox enzymes. siNOX4 was able to decrease the level of ROS production. Myofibroblast differentiation and the production of collagen in NPDFs were prevented by inhibition of ROS generation with diphenyliodonium, N-acetylcysteine, ebselen, and siNox4. Conclusions: This study showed that NOX4 and ROS have a role in myofibroblast differentiation and collagen production of TGF-β1-induced NPDFs and that these processes are inhibited by the elimination of ROS.

AB - Background: Myofibroblasts are detected in nasal polyps and are involved in nasal polyp formation by inducing extracellular matrix accumulation. Reactive oxygen species (ROS) are released during the differentiation of fibroblasts to myofibroblasts. The purpose of this study was to investigate ROS production and nicotinamide adenine dinucleotide phosphate oxidase (NOX) expression in nasal polyp-derived fibroblasts (NPDFs) and to evaluate whether ROS from NOX mediates transforming growth factor (TGF)-β1-induced production of alpha smooth-muscle actin (α-SMA) and collagen production. Methods: NPDFs were incubated and treated with TGF-β1. The mRNA expression of NOXs, α-SMA, and collagen type I and IV was determined by reverse transcription-polymerase chain reaction, and the expression of α-SMA protein was determined by immunofluorescence microscopy. The amount of total soluble collagen production was analyzed by the SirCol assay. The ROS generation of cells was investigated using the 2′,7′-dichlorfluorescein- diacetate. The fluorescence was captured by fluorescent microscope and measured using a fluorometer. Results: Stimulation with TGF-β1 increased ROS production by NPDFs compared with NPDFs not treated with TGF-β1. Stimulation with TGF-β1 increased the expression of NOX4 mRNA most potently among various Nox enzymes. siNOX4 was able to decrease the level of ROS production. Myofibroblast differentiation and the production of collagen in NPDFs were prevented by inhibition of ROS generation with diphenyliodonium, N-acetylcysteine, ebselen, and siNox4. Conclusions: This study showed that NOX4 and ROS have a role in myofibroblast differentiation and collagen production of TGF-β1-induced NPDFs and that these processes are inhibited by the elimination of ROS.

KW - Alpha-smooth-muscle actin

KW - Myofibroblast

KW - Nasal polyps

KW - Nicotinamide adenine dinucleotide phosphate oxidase

KW - Reactive oxygen species

KW - Transforming growth factor-beta1

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U2 - 10.1159/000337460

DO - 10.1159/000337460

M3 - Article

VL - 159

SP - 278

EP - 286

JO - International Archives of Allergy and Immunology

JF - International Archives of Allergy and Immunology

SN - 1018-2438

IS - 3

ER -