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

Yan Liu, Hong Xu, Yucong Geng, Dingjie Xu, Lijuan Zhang, Yi Yang, Zhongqiu Wei, Bonan Zhang, Shifeng Li, Xuemin Gao, Ruimin Wang, Xianghong Zhang, Darrell W Brann, Fang Yang

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

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) 10-7mol/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.

Original languageEnglish (US)
Article number38
JournalRespiratory Research
Volume18
Issue number1
DOIs
StatePublished - Feb 21 2017

Fingerprint

CREB-Binding Protein
Silicosis
Myofibroblasts
Pulmonary Fibrosis
Cyclic AMP-Dependent Protein Kinases
Cyclic AMP Response Element-Binding Protein
Angiotensin II
Extracellular Matrix
Fibroblasts
Staining and Labeling
Lung
Proteins
Therapeutics
Hematoxylin
Eosine Yellowish-(YS)
Immunoenzyme Techniques
Immunoprecipitation
Protein Binding
Silicon Dioxide
Fluorescent Antibody Technique

Keywords

  • CAMP
  • CREB
  • Myofibroblast
  • PKA
  • Silicosis
  • Smad

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

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 W; Yang, Fang.

In: Respiratory Research, Vol. 18, No. 1, 38, 21.02.2017.

Research output: Contribution to journalArticle

Liu, Y, Xu, H, Geng, Y, Xu, D, Zhang, L, Yang, Y, Wei, Z, Zhang, B, Li, S, Gao, X, Wang, R, Zhang, X, Brann, DW & Yang, F 2017, 'Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis', Respiratory Research, vol. 18, no. 1, 38. https://doi.org/10.1186/s12931-017-0523-z
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 W ; Yang, Fang. / Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis. In: Respiratory Research. 2017 ; Vol. 18, No. 1.
@article{1f45d8e52a8343b88b91dc277b13b6a7,
title = "Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis",
abstract = "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) 10-7mol/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.",
keywords = "CAMP, CREB, Myofibroblast, PKA, Silicosis, Smad",
author = "Yan Liu and Hong Xu and Yucong Geng and Dingjie Xu and Lijuan Zhang and Yi Yang and Zhongqiu Wei and Bonan Zhang and Shifeng Li and Xuemin Gao and Ruimin Wang and Xianghong Zhang and Brann, {Darrell W} and Fang Yang",
year = "2017",
month = "2",
day = "21",
doi = "10.1186/s12931-017-0523-z",
language = "English (US)",
volume = "18",
journal = "Respiratory Research",
issn = "1465-9921",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Dibutyryl-cAMP attenuates pulmonary fibrosis by blocking myofibroblast differentiation via PKA/CREB/CBP signaling in rats with silicosis

AU - Liu, Yan

AU - Xu, Hong

AU - Geng, Yucong

AU - Xu, Dingjie

AU - Zhang, Lijuan

AU - Yang, Yi

AU - Wei, Zhongqiu

AU - Zhang, Bonan

AU - Li, Shifeng

AU - Gao, Xuemin

AU - Wang, Ruimin

AU - Zhang, Xianghong

AU - Brann, Darrell W

AU - Yang, Fang

PY - 2017/2/21

Y1 - 2017/2/21

N2 - 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) 10-7mol/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.

AB - 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) 10-7mol/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.

KW - CAMP

KW - CREB

KW - Myofibroblast

KW - PKA

KW - Silicosis

KW - Smad

UR - http://www.scopus.com/inward/record.url?scp=85014421994&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85014421994&partnerID=8YFLogxK

U2 - 10.1186/s12931-017-0523-z

DO - 10.1186/s12931-017-0523-z

M3 - Article

C2 - 28222740

AN - SCOPUS:85014421994

VL - 18

JO - Respiratory Research

JF - Respiratory Research

SN - 1465-9921

IS - 1

M1 - 38

ER -