Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis

BACKGROUND: Myofibroblasts play a major role in the synthesis of extracellular matrix (ECM) and the stimulation of these cells is thought to play an important role in the development of silicosis. The present study was undertaken to investigate the anti-fibrotic effects of dibutyryl-cAMP (db-cAMP) o...

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Autores principales: Liu, Yan, Xu, Hong, Geng, Yucong, Xu, Dingjie, Zhang, Lijuan, Yang, Yi, Wei, Zhongqiu, Zhang, Bonan, Li, Shifeng, Gao, Xuemin, Wang, Ruimin, Zhang, Xianghong, Brann, Darrell, Yang, Fang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5320641/
https://www.ncbi.nlm.nih.gov/pubmed/28222740
http://dx.doi.org/10.1186/s12931-017-0523-z
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author Liu, Yan
Xu, Hong
Geng, Yucong
Xu, Dingjie
Zhang, Lijuan
Yang, Yi
Wei, Zhongqiu
Zhang, Bonan
Li, Shifeng
Gao, Xuemin
Wang, Ruimin
Zhang, Xianghong
Brann, Darrell
Yang, Fang
author_facet Liu, Yan
Xu, Hong
Geng, Yucong
Xu, Dingjie
Zhang, Lijuan
Yang, Yi
Wei, Zhongqiu
Zhang, Bonan
Li, Shifeng
Gao, Xuemin
Wang, Ruimin
Zhang, Xianghong
Brann, Darrell
Yang, Fang
author_sort Liu, Yan
collection PubMed
description BACKGROUND: Myofibroblasts play a major role in the synthesis of extracellular matrix (ECM) and the stimulation of these cells is thought to play an important role in the development of silicosis. The present study was undertaken to investigate the anti-fibrotic effects of dibutyryl-cAMP (db-cAMP) on rats induced by silica. METHODS: A HOPE MED 8050 exposure control apparatus was used to create the silicosis model. Rats were randomly divided into 4 groups: 1)controls for 16 w; 2)silicosis for 16 w; 3)db-cAMP pre-treatment; 4) db-cAMP post-treatment. Rat pulmonary fibroblasts were cultured in vitro and divided into 4 groups as follows: 1) controls; 2) 10(−7)mol/L angiotensin II (Ang II); 3) Ang II +10(−4) mol/L db-cAMP; and 4) Ang II + db-cAMP+ 10(−6) mol/L H89. Hematoxylin-eosin (HE), Van Gieson staining and immunohistochemistry (IHC) were performed to observe the histomorphology of lung tissue. The levels of cAMP were detected by enzyme immunoassay. Double-labeling for α-SMA with Gαi3, protein kinase A (PKA), phosphorylated cAMP-response element-binding protein (p-CREB), and p-Smad2/3 was identified by immunofluorescence staining. Protein levels were detected by Western blot analysis. The interaction between CREB-binding protein (CBP) and Smad2/3 and p-CREB were measured by co-immunoprecipitation (Co-IP). RESULTS: Db-cAMP treatment reduced the number and size of silicosis nodules, inhibited myofibroblast differentiation, and extracellular matrix deposition in vitro and in vivo. In addition, db-cAMP regulated Gαs protein and inhibited expression of Gαi protein, which increased endogenous cAMP. Db-cAMP increased phosphorylated cAMP-response element-binding protein (p-CREB) via protein kinase A (PKA) signaling, and decreased nuclear p-Smad2/3 binding with CREB binding protein (CBP), which reduced activation of p-Smads in fibroblasts induced by Ang II. CONCLUSIONS: This study showed an anti-silicotic effect of db-cAMP that was mediated via PKA/p-CREB/CBP signaling. Furthermore, the findings offer novel insight into the potential use of cAMP signaling for therapeutic strategies to treat silicosis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12931-017-0523-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-53206412017-02-24 Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis Liu, Yan Xu, Hong Geng, Yucong Xu, Dingjie Zhang, Lijuan Yang, Yi Wei, Zhongqiu Zhang, Bonan Li, Shifeng Gao, Xuemin Wang, Ruimin Zhang, Xianghong Brann, Darrell Yang, Fang Respir Res Research BACKGROUND: Myofibroblasts play a major role in the synthesis of extracellular matrix (ECM) and the stimulation of these cells is thought to play an important role in the development of silicosis. The present study was undertaken to investigate the anti-fibrotic effects of dibutyryl-cAMP (db-cAMP) on rats induced by silica. METHODS: A HOPE MED 8050 exposure control apparatus was used to create the silicosis model. Rats were randomly divided into 4 groups: 1)controls for 16 w; 2)silicosis for 16 w; 3)db-cAMP pre-treatment; 4) db-cAMP post-treatment. Rat pulmonary fibroblasts were cultured in vitro and divided into 4 groups as follows: 1) controls; 2) 10(−7)mol/L angiotensin II (Ang II); 3) Ang II +10(−4) mol/L db-cAMP; and 4) Ang II + db-cAMP+ 10(−6) mol/L H89. Hematoxylin-eosin (HE), Van Gieson staining and immunohistochemistry (IHC) were performed to observe the histomorphology of lung tissue. The levels of cAMP were detected by enzyme immunoassay. Double-labeling for α-SMA with Gαi3, protein kinase A (PKA), phosphorylated cAMP-response element-binding protein (p-CREB), and p-Smad2/3 was identified by immunofluorescence staining. Protein levels were detected by Western blot analysis. The interaction between CREB-binding protein (CBP) and Smad2/3 and p-CREB were measured by co-immunoprecipitation (Co-IP). RESULTS: Db-cAMP treatment reduced the number and size of silicosis nodules, inhibited myofibroblast differentiation, and extracellular matrix deposition in vitro and in vivo. In addition, db-cAMP regulated Gαs protein and inhibited expression of Gαi protein, which increased endogenous cAMP. Db-cAMP increased phosphorylated cAMP-response element-binding protein (p-CREB) via protein kinase A (PKA) signaling, and decreased nuclear p-Smad2/3 binding with CREB binding protein (CBP), which reduced activation of p-Smads in fibroblasts induced by Ang II. CONCLUSIONS: This study showed an anti-silicotic effect of db-cAMP that was mediated via PKA/p-CREB/CBP signaling. Furthermore, the findings offer novel insight into the potential use of cAMP signaling for therapeutic strategies to treat silicosis. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12931-017-0523-z) contains supplementary material, which is available to authorized users. BioMed Central 2017-02-21 2017 /pmc/articles/PMC5320641/ /pubmed/28222740 http://dx.doi.org/10.1186/s12931-017-0523-z Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Liu, Yan
Xu, Hong
Geng, Yucong
Xu, Dingjie
Zhang, Lijuan
Yang, Yi
Wei, Zhongqiu
Zhang, Bonan
Li, Shifeng
Gao, Xuemin
Wang, Ruimin
Zhang, Xianghong
Brann, Darrell
Yang, Fang
Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title_full Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title_fullStr Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title_full_unstemmed Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title_short Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis
title_sort dibutyryl-camp attenuates pulmonary fibrosis by blocking myofibroblast differentiation via pka/creb/cbp signaling in rats with silicosis
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5320641/
https://www.ncbi.nlm.nih.gov/pubmed/28222740
http://dx.doi.org/10.1186/s12931-017-0523-z
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