Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes

Yukiko Namba, Shinsaku Togo, Miniwan Tulafu, Kotaro Kadoya, Kumi Yoneda Nagahama, Hikari Taka, Naoko Kaga, Akira Orimo, Xiang-de Liu, Kazuhisa Takahashi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Airway fibrosis is one of the pathological features of chronic obstructive pulmonary disease (COPD), and recent studies revealed that acetylcholine plays an important role in the development of airway remodeling by stimulating proliferation and collagen synthesis of lung fibroblasts. This study was designed to examine the effects of a long-acting muscarinic receptor antagonist (LAMA) glycopyrronium and a long-acting β2 adrenergic receptor agonist (LABA) indacaterol on acetylcholine-mediated fibrotic responses in lung fibroblasts. Methods: After carbachol (CCh) or transforming growth factor-β1 (TGF-β1) exposure, the response to glycopyrronium and indacaterol was determined in vitro in fibroblasts isolated from mild-to-moderate COPD lung tissue. The ability of fibroblasts to mediate the contraction of collagen gels was assessed. The expression of α-smooth muscle actin (α-SMA) and the phosphorylation of extracellular-signal-regulated kinase 5 (ERK5) were determined by immunoblot. TGF-β1 was quantified by ELISA and acetylcholine was quantified by liquid chromatography tandem-mass spectrometry. Results: CCh stimulated fibroblast-mediated collagen gel contraction and α-SMA expression and TGF-β1 release by fibroblasts. Blockade of autocrine TGF-β1 attenuated CCh-mediated fibrotic responses, while TGF-β1 did not stimulate acetylcholine release. Glycopyrronium plus indacaterol significantly attenuated CCh- and TGF-β1-mediated fibrotic responses through inhibition of ERK5 phosphorylation. Notably, the magnitudes of CCh- and TGF-β1-stimulated gel contraction, CCh-induced TGF-β1 release, and ERK5 phosphorylation were greater in fibroblasts isolated from COPD subjects than in those from non-smokers. Conclusions: CCh induced TGF-β1 self-sustaining signaling loops by potentiating ERK5 signaling and promoted myofibroblast activity. This autocrine signaling mechanism may be an attractive therapeutic target to block the fibrotic response, which was modulated by the combination of glycopyrronium and indacaterol.

Original languageEnglish (US)
Article number46
JournalRespiratory Research
Volume18
Issue number1
DOIs
StatePublished - Mar 11 2017

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Mitogen-Activated Protein Kinase 7
Carbachol
Transforming Growth Factors
Fibroblasts
Acetylcholine
Chronic Obstructive Pulmonary Disease
Collagen
Gels
Phosphorylation
Lung
Autocrine Communication
Airway Remodeling
indacaterol
Adrenergic Agonists
Muscarinic Antagonists
Myofibroblasts
Muscarinic Receptors
Tandem Mass Spectrometry
Liquid Chromatography
Smooth Muscle

Keywords

  • Acetylcholine
  • Extracellular-signal-regulated kinase 5 (ERK5)
  • Long-acting muscarinic receptor antagonist
  • Long-acting β2-adrenergic receptor agonist
  • Transforming growth factor-β1

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Namba, Y., Togo, S., Tulafu, M., Kadoya, K., Nagahama, K. Y., Taka, H., ... Takahashi, K. (2017). Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes. Respiratory Research, 18(1), [46]. https://doi.org/10.1186/s12931-017-0529-6

Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes. / Namba, Yukiko; Togo, Shinsaku; Tulafu, Miniwan; Kadoya, Kotaro; Nagahama, Kumi Yoneda; Taka, Hikari; Kaga, Naoko; Orimo, Akira; Liu, Xiang-de; Takahashi, Kazuhisa.

In: Respiratory Research, Vol. 18, No. 1, 46, 11.03.2017.

Research output: Contribution to journalArticle

Namba, Y, Togo, S, Tulafu, M, Kadoya, K, Nagahama, KY, Taka, H, Kaga, N, Orimo, A, Liu, X & Takahashi, K 2017, 'Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes', Respiratory Research, vol. 18, no. 1, 46. https://doi.org/10.1186/s12931-017-0529-6
Namba, Yukiko ; Togo, Shinsaku ; Tulafu, Miniwan ; Kadoya, Kotaro ; Nagahama, Kumi Yoneda ; Taka, Hikari ; Kaga, Naoko ; Orimo, Akira ; Liu, Xiang-de ; Takahashi, Kazuhisa. / Combination of glycopyrronium and indacaterol inhibits carbachol-induced ERK5 signal in fibrotic processes. In: Respiratory Research. 2017 ; Vol. 18, No. 1.
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abstract = "Background: Airway fibrosis is one of the pathological features of chronic obstructive pulmonary disease (COPD), and recent studies revealed that acetylcholine plays an important role in the development of airway remodeling by stimulating proliferation and collagen synthesis of lung fibroblasts. This study was designed to examine the effects of a long-acting muscarinic receptor antagonist (LAMA) glycopyrronium and a long-acting β2 adrenergic receptor agonist (LABA) indacaterol on acetylcholine-mediated fibrotic responses in lung fibroblasts. Methods: After carbachol (CCh) or transforming growth factor-β1 (TGF-β1) exposure, the response to glycopyrronium and indacaterol was determined in vitro in fibroblasts isolated from mild-to-moderate COPD lung tissue. The ability of fibroblasts to mediate the contraction of collagen gels was assessed. The expression of α-smooth muscle actin (α-SMA) and the phosphorylation of extracellular-signal-regulated kinase 5 (ERK5) were determined by immunoblot. TGF-β1 was quantified by ELISA and acetylcholine was quantified by liquid chromatography tandem-mass spectrometry. Results: CCh stimulated fibroblast-mediated collagen gel contraction and α-SMA expression and TGF-β1 release by fibroblasts. Blockade of autocrine TGF-β1 attenuated CCh-mediated fibrotic responses, while TGF-β1 did not stimulate acetylcholine release. Glycopyrronium plus indacaterol significantly attenuated CCh- and TGF-β1-mediated fibrotic responses through inhibition of ERK5 phosphorylation. Notably, the magnitudes of CCh- and TGF-β1-stimulated gel contraction, CCh-induced TGF-β1 release, and ERK5 phosphorylation were greater in fibroblasts isolated from COPD subjects than in those from non-smokers. Conclusions: CCh induced TGF-β1 self-sustaining signaling loops by potentiating ERK5 signaling and promoted myofibroblast activity. This autocrine signaling mechanism may be an attractive therapeutic target to block the fibrotic response, which was modulated by the combination of glycopyrronium and indacaterol.",
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AU - Kadoya, Kotaro

AU - Nagahama, Kumi Yoneda

AU - Taka, Hikari

AU - Kaga, Naoko

AU - Orimo, Akira

AU - Liu, Xiang-de

AU - Takahashi, Kazuhisa

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N2 - Background: Airway fibrosis is one of the pathological features of chronic obstructive pulmonary disease (COPD), and recent studies revealed that acetylcholine plays an important role in the development of airway remodeling by stimulating proliferation and collagen synthesis of lung fibroblasts. This study was designed to examine the effects of a long-acting muscarinic receptor antagonist (LAMA) glycopyrronium and a long-acting β2 adrenergic receptor agonist (LABA) indacaterol on acetylcholine-mediated fibrotic responses in lung fibroblasts. Methods: After carbachol (CCh) or transforming growth factor-β1 (TGF-β1) exposure, the response to glycopyrronium and indacaterol was determined in vitro in fibroblasts isolated from mild-to-moderate COPD lung tissue. The ability of fibroblasts to mediate the contraction of collagen gels was assessed. The expression of α-smooth muscle actin (α-SMA) and the phosphorylation of extracellular-signal-regulated kinase 5 (ERK5) were determined by immunoblot. TGF-β1 was quantified by ELISA and acetylcholine was quantified by liquid chromatography tandem-mass spectrometry. Results: CCh stimulated fibroblast-mediated collagen gel contraction and α-SMA expression and TGF-β1 release by fibroblasts. Blockade of autocrine TGF-β1 attenuated CCh-mediated fibrotic responses, while TGF-β1 did not stimulate acetylcholine release. Glycopyrronium plus indacaterol significantly attenuated CCh- and TGF-β1-mediated fibrotic responses through inhibition of ERK5 phosphorylation. Notably, the magnitudes of CCh- and TGF-β1-stimulated gel contraction, CCh-induced TGF-β1 release, and ERK5 phosphorylation were greater in fibroblasts isolated from COPD subjects than in those from non-smokers. Conclusions: CCh induced TGF-β1 self-sustaining signaling loops by potentiating ERK5 signaling and promoted myofibroblast activity. This autocrine signaling mechanism may be an attractive therapeutic target to block the fibrotic response, which was modulated by the combination of glycopyrronium and indacaterol.

AB - Background: Airway fibrosis is one of the pathological features of chronic obstructive pulmonary disease (COPD), and recent studies revealed that acetylcholine plays an important role in the development of airway remodeling by stimulating proliferation and collagen synthesis of lung fibroblasts. This study was designed to examine the effects of a long-acting muscarinic receptor antagonist (LAMA) glycopyrronium and a long-acting β2 adrenergic receptor agonist (LABA) indacaterol on acetylcholine-mediated fibrotic responses in lung fibroblasts. Methods: After carbachol (CCh) or transforming growth factor-β1 (TGF-β1) exposure, the response to glycopyrronium and indacaterol was determined in vitro in fibroblasts isolated from mild-to-moderate COPD lung tissue. The ability of fibroblasts to mediate the contraction of collagen gels was assessed. The expression of α-smooth muscle actin (α-SMA) and the phosphorylation of extracellular-signal-regulated kinase 5 (ERK5) were determined by immunoblot. TGF-β1 was quantified by ELISA and acetylcholine was quantified by liquid chromatography tandem-mass spectrometry. Results: CCh stimulated fibroblast-mediated collagen gel contraction and α-SMA expression and TGF-β1 release by fibroblasts. Blockade of autocrine TGF-β1 attenuated CCh-mediated fibrotic responses, while TGF-β1 did not stimulate acetylcholine release. Glycopyrronium plus indacaterol significantly attenuated CCh- and TGF-β1-mediated fibrotic responses through inhibition of ERK5 phosphorylation. Notably, the magnitudes of CCh- and TGF-β1-stimulated gel contraction, CCh-induced TGF-β1 release, and ERK5 phosphorylation were greater in fibroblasts isolated from COPD subjects than in those from non-smokers. Conclusions: CCh induced TGF-β1 self-sustaining signaling loops by potentiating ERK5 signaling and promoted myofibroblast activity. This autocrine signaling mechanism may be an attractive therapeutic target to block the fibrotic response, which was modulated by the combination of glycopyrronium and indacaterol.

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KW - Extracellular-signal-regulated kinase 5 (ERK5)

KW - Long-acting muscarinic receptor antagonist

KW - Long-acting β2-adrenergic receptor agonist

KW - Transforming growth factor-β1

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