The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1

Sang Hoon Lee, Ju Hye Shin, Mi Hwa Shin, Young Sam Kim, Kyung Soo Chung, Joo Han Song, Song Yee Kim, Eun Young Kim, Ji Ye Jung, youngae kang, Joon Chang, Moo Suk Park

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Abstract

Background: Transforming growth factor β (TGF-β), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-β1. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-β1 and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-β1 and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-β1 prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-β1 stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-β1 failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-β1-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-β1 and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-β1 in vitro , and RA also decreased the expression of TGF-β1 at 1 and 3 weeks in vivo . Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-β1/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-β1-induced lung injury and fibrosis.

Original languageEnglish
Pages (from-to)42-52
Number of pages11
JournalTuberculosis and Respiratory Diseases
Volume82
Issue number1
DOIs
Publication statusPublished - 2019 Jan 1

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Transforming Growth Factors
Protein Kinase Inhibitors
Tretinoin
Mitogen-Activated Protein Kinases
Phosphorylation
Lung Injury
Mitogen-Activated Protein Kinase Kinases
Bleomycin
p38 Mitogen-Activated Protein Kinases
Fibroblasts
Epithelial Cells
Cell Differentiation
Fibrosis
Animal Models
Western Blotting
Cell Proliferation

All Science Journal Classification (ASJC) codes

  • Pulmonary and Respiratory Medicine
  • Infectious Diseases

Cite this

Lee, Sang Hoon ; Shin, Ju Hye ; Shin, Mi Hwa ; Kim, Young Sam ; Chung, Kyung Soo ; Song, Joo Han ; Kim, Song Yee ; Kim, Eun Young ; Jung, Ji Ye ; kang, youngae ; Chang, Joon ; Park, Moo Suk. / The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1. In: Tuberculosis and Respiratory Diseases. 2019 ; Vol. 82, No. 1. pp. 42-52.
@article{5e92a815889a4053abf5dfd0f3612e34,
title = "The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1",
abstract = "Background: Transforming growth factor β (TGF-β), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-β1. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-β1 and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-β1 and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-β1 prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-β1 stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-β1 failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-β1-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-β1 and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-β1 in vitro , and RA also decreased the expression of TGF-β1 at 1 and 3 weeks in vivo . Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-β1/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-β1-induced lung injury and fibrosis.",
author = "Lee, {Sang Hoon} and Shin, {Ju Hye} and Shin, {Mi Hwa} and Kim, {Young Sam} and Chung, {Kyung Soo} and Song, {Joo Han} and Kim, {Song Yee} and Kim, {Eun Young} and Jung, {Ji Ye} and youngae kang and Joon Chang and Park, {Moo Suk}",
year = "2019",
month = "1",
day = "1",
doi = "10.4046/trd.2017.0111",
language = "English",
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Lee, SH, Shin, JH, Shin, MH, Kim, YS, Chung, KS, Song, JH, Kim, SY, Kim, EY, Jung, JY, kang, Y, Chang, J & Park, MS 2019, 'The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1', Tuberculosis and Respiratory Diseases, vol. 82, no. 1, pp. 42-52. https://doi.org/10.4046/trd.2017.0111

The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1. / Lee, Sang Hoon; Shin, Ju Hye; Shin, Mi Hwa; Kim, Young Sam; Chung, Kyung Soo; Song, Joo Han; Kim, Song Yee; Kim, Eun Young; Jung, Ji Ye; kang, youngae; Chang, Joon; Park, Moo Suk.

In: Tuberculosis and Respiratory Diseases, Vol. 82, No. 1, 01.01.2019, p. 42-52.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The effects of retinoic acid and MAPK inhibitors on phosphorylation of Smad2/3 induced by transforming growth factor β1

AU - Lee, Sang Hoon

AU - Shin, Ju Hye

AU - Shin, Mi Hwa

AU - Kim, Young Sam

AU - Chung, Kyung Soo

AU - Song, Joo Han

AU - Kim, Song Yee

AU - Kim, Eun Young

AU - Jung, Ji Ye

AU - kang, youngae

AU - Chang, Joon

AU - Park, Moo Suk

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Background: Transforming growth factor β (TGF-β), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-β1. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-β1 and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-β1 and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-β1 prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-β1 stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-β1 failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-β1-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-β1 and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-β1 in vitro , and RA also decreased the expression of TGF-β1 at 1 and 3 weeks in vivo . Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-β1/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-β1-induced lung injury and fibrosis.

AB - Background: Transforming growth factor β (TGF-β), retinoic acid (RA), p38 mitogen-activated protein kinase (MAPK), and MEK signaling play critical roles in cell differentiation, proliferation, and apoptosis. We investigated the effect of RA and the role of these signaling molecules on the phosphorylation of Smad2/3 (p-Smad2/3) induced by TGF-β1. Methods: A549 epithelial cells and CCD-11Lu fibroblasts were incubated and stimulated with or without all-trans RA (ATRA) and TGF-β1 and with MAPK or MEK inhibitors. The levels of p-Smad2/3 were analyzed by western blotting. For animal models, we studied three experimental mouse groups: control, bleomycin, and bleomycin+ATRA group. Changes in histopathology, lung injury score, and levels of TGF-β1 and Smad3 were evaluated at 1 and 3 weeks. Results: When A549 cells were pre-stimulated with TGF-β1 prior to RA treatment, RA completely inhibited the p-Smad2/3. However, when A549 cells were pre-treated with RA prior to TGF-β1 stimulation, RA did not completely suppress the p-Smad2/3. When A549 cells were pre-treated with MAPK inhibitor, TGF-β1 failed to phosphorylate Smad2/3. In fibroblasts, p38 MAPK inhibitor suppressed TGF-β1-induced p-Smad2. In a bleomycin-induced lung injury mouse model, RA decreased the expression of TGF-β1 and Smad3 at 1 and 3 weeks. Conclusion: RA had inhibitory effects on the phosphorylation of Smad induced by TGF-β1 in vitro , and RA also decreased the expression of TGF-β1 at 1 and 3 weeks in vivo . Furthermore, pre-treatment with a MAPK inhibitor showed a preventative effect on TGF-β1/Smad phosphorylation in epithelial cells. As a result, a combination of RA and MAPK inhibitors may suppress the TGF-β1-induced lung injury and fibrosis.

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