Knockout of endothelin type B receptor signaling attenuates bleomycin-induced skin sclerosis in mice

Kengo Akashi, Jun Saegusa, Sho Sendo, Keisuke Nishimura, Takuya Okano, Keiko Yagi, Masashi Yanagisawa, Noriaki Emoto, Akio Morinobu

Research output: Contribution to journalArticle

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Abstract

Background: Endothelin-1 (ET-1) is important in the pathogenesis of systemic sclerosis (SSc). ET-1 binds two receptors, endothelin type A (ETA) and endothelin type B (ETB). Dual ETA/ETB receptor antagonists and a selective ETA receptor antagonist are used clinically to treat SSc, and the effect of these antagonists on fibroblast activation has been described. However, the role of ETB receptor signaling in fibrogenesis is less clear. This study was conducted to evaluate the profibrotic function of ETB receptor signaling in a murine model of bleomycin (BLM)-induced scleroderma. Methods: We used ETB receptor-knockout (ETBKO) mice, which are genetically rescued from lethal intestinal aganglionosis by an ETB receptor transgene driven by the human dopamine β-hydroxylase (DβH)-gene promoter, and wild-type mice with DβH-ETB (WT). BLM or phosphate-buffered saline (PBS) was administered subcutaneously by osmotic minipump, and skin fibrosis was assessed by dermal thickness, subcutaneous fat atrophy, and myofibroblast count in the dermis. Dermal fibroblasts isolated from ETBKO and WT mice were cultured in vitro, stimulated with BLM or ET-1, and the expression of profibrotic genes was compared by quantitative PCR. Results: Dermal thickness, subcutaneous fat atrophy, and myofibroblast counts in the dermis were significantly reduced in ETBKO mice compared to WT mice, after BLM treatment. Compared with wild-type, dermal fibroblasts isolated from ETBKO mice showed lower gene expressions of α-smooth muscle actin and collagen 1α1 in response to BLM or ET-1 stimulation in vitro. Conclusions: ET-1-ETB receptor signaling is involved in skin sclerosis and in collagen synthesis by dermal fibroblasts.

Original languageEnglish (US)
Article number113
JournalArthritis Research and Therapy
Volume18
Issue number1
DOIs
StatePublished - May 21 2016

Fingerprint

Endothelin B Receptors
Bleomycin
Sclerosis
Endothelin-1
Skin
Knockout Mice
Endothelins
Fibroblasts
Myofibroblasts
Systemic Scleroderma
Subcutaneous Fat
Dermis
Mixed Function Oxygenases
Atrophy
Dopamine
Collagen
Gene Expression
Endothelin A Receptors
Transgenes
Smooth Muscle

Keywords

  • Dermal fibroblast
  • Endothelin type B receptor
  • Systemic sclerosis

ASJC Scopus subject areas

  • Rheumatology
  • Immunology
  • Immunology and Allergy

Cite this

Akashi, K., Saegusa, J., Sendo, S., Nishimura, K., Okano, T., Yagi, K., ... Morinobu, A. (2016). Knockout of endothelin type B receptor signaling attenuates bleomycin-induced skin sclerosis in mice. Arthritis Research and Therapy, 18(1), [113]. https://doi.org/10.1186/s13075-016-1011-4

Knockout of endothelin type B receptor signaling attenuates bleomycin-induced skin sclerosis in mice. / Akashi, Kengo; Saegusa, Jun; Sendo, Sho; Nishimura, Keisuke; Okano, Takuya; Yagi, Keiko; Yanagisawa, Masashi; Emoto, Noriaki; Morinobu, Akio.

In: Arthritis Research and Therapy, Vol. 18, No. 1, 113, 21.05.2016.

Research output: Contribution to journalArticle

Akashi, Kengo ; Saegusa, Jun ; Sendo, Sho ; Nishimura, Keisuke ; Okano, Takuya ; Yagi, Keiko ; Yanagisawa, Masashi ; Emoto, Noriaki ; Morinobu, Akio. / Knockout of endothelin type B receptor signaling attenuates bleomycin-induced skin sclerosis in mice. In: Arthritis Research and Therapy. 2016 ; Vol. 18, No. 1.
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abstract = "Background: Endothelin-1 (ET-1) is important in the pathogenesis of systemic sclerosis (SSc). ET-1 binds two receptors, endothelin type A (ETA) and endothelin type B (ETB). Dual ETA/ETB receptor antagonists and a selective ETA receptor antagonist are used clinically to treat SSc, and the effect of these antagonists on fibroblast activation has been described. However, the role of ETB receptor signaling in fibrogenesis is less clear. This study was conducted to evaluate the profibrotic function of ETB receptor signaling in a murine model of bleomycin (BLM)-induced scleroderma. Methods: We used ETB receptor-knockout (ETBKO) mice, which are genetically rescued from lethal intestinal aganglionosis by an ETB receptor transgene driven by the human dopamine β-hydroxylase (DβH)-gene promoter, and wild-type mice with DβH-ETB (WT). BLM or phosphate-buffered saline (PBS) was administered subcutaneously by osmotic minipump, and skin fibrosis was assessed by dermal thickness, subcutaneous fat atrophy, and myofibroblast count in the dermis. Dermal fibroblasts isolated from ETBKO and WT mice were cultured in vitro, stimulated with BLM or ET-1, and the expression of profibrotic genes was compared by quantitative PCR. Results: Dermal thickness, subcutaneous fat atrophy, and myofibroblast counts in the dermis were significantly reduced in ETBKO mice compared to WT mice, after BLM treatment. Compared with wild-type, dermal fibroblasts isolated from ETBKO mice showed lower gene expressions of α-smooth muscle actin and collagen 1α1 in response to BLM or ET-1 stimulation in vitro. Conclusions: ET-1-ETB receptor signaling is involved in skin sclerosis and in collagen synthesis by dermal fibroblasts.",
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AU - Sendo, Sho

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AU - Okano, Takuya

AU - Yagi, Keiko

AU - Yanagisawa, Masashi

AU - Emoto, Noriaki

AU - Morinobu, Akio

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AB - Background: Endothelin-1 (ET-1) is important in the pathogenesis of systemic sclerosis (SSc). ET-1 binds two receptors, endothelin type A (ETA) and endothelin type B (ETB). Dual ETA/ETB receptor antagonists and a selective ETA receptor antagonist are used clinically to treat SSc, and the effect of these antagonists on fibroblast activation has been described. However, the role of ETB receptor signaling in fibrogenesis is less clear. This study was conducted to evaluate the profibrotic function of ETB receptor signaling in a murine model of bleomycin (BLM)-induced scleroderma. Methods: We used ETB receptor-knockout (ETBKO) mice, which are genetically rescued from lethal intestinal aganglionosis by an ETB receptor transgene driven by the human dopamine β-hydroxylase (DβH)-gene promoter, and wild-type mice with DβH-ETB (WT). BLM or phosphate-buffered saline (PBS) was administered subcutaneously by osmotic minipump, and skin fibrosis was assessed by dermal thickness, subcutaneous fat atrophy, and myofibroblast count in the dermis. Dermal fibroblasts isolated from ETBKO and WT mice were cultured in vitro, stimulated with BLM or ET-1, and the expression of profibrotic genes was compared by quantitative PCR. Results: Dermal thickness, subcutaneous fat atrophy, and myofibroblast counts in the dermis were significantly reduced in ETBKO mice compared to WT mice, after BLM treatment. Compared with wild-type, dermal fibroblasts isolated from ETBKO mice showed lower gene expressions of α-smooth muscle actin and collagen 1α1 in response to BLM or ET-1 stimulation in vitro. Conclusions: ET-1-ETB receptor signaling is involved in skin sclerosis and in collagen synthesis by dermal fibroblasts.

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