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Periostin facilitates skin sclerosis via PI3K/Akt dependent mechanism in a mouse model of scleroderma.

Yang L, Serada S, Fujimoto M, Terao M, Kotobuki Y, Kitaba S, Matsui S, Kudo A, Naka T, Murota H, Katayama I - PLoS ONE (2012)

Bottom Line: Periostin, a novel matricellular protein, is recently reported to play a crucial role in tissue remodeling and is highly expressed under fibrotic conditions.To determine the downstream pathway involved in periostin signaling, receptor neutralizing antibody and signal transduction inhibitors were used in vitro.Furthermore, recombinant mouse periostin directly induced Col1α1 expression in vitro, and this effect was inhibited by blocking the αv integrin-mediated PI3K/Akt signaling either with anti-αv functional blocking antibody or with the PI3K/Akt kinase inhibitor LY294002.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Osaka University Graduate School of Medicine, Osaka, Japan.

ABSTRACT

Objective: Periostin, a novel matricellular protein, is recently reported to play a crucial role in tissue remodeling and is highly expressed under fibrotic conditions. This study was undertaken to assess the role of periostin in scleroderma.

Methods: Using skin from patients and healthy donors, the expression of periostin was assessed by immunohistochemistry and immunoblotting analyses. Furthermore, we investigated periostin(-/-) (PN(-/-)) and wild-type (WT) mice to elucidate the role of periostin in scleroderma. To induce murine cutaneous sclerosis, mice were subcutaneously injected with bleomycin, while untreated control groups were injected with phosphate-buffered saline. Bleomycin-induced fibrotic changes were compared in PN(-/-) and WT mice by histological analysis as well as by measurements of profibrotic cytokine and extracellular matrix protein expression levels in vivo and in vitro. To determine the downstream pathway involved in periostin signaling, receptor neutralizing antibody and signal transduction inhibitors were used in vitro.

Results: Elevated expression of periostin was observed in the lesional skin of patients with scleroderma compared with healthy donors. Although WT mice showed marked cutaneous sclerosis with increased expression of periostin and increased numbers of myofibroblasts after bleomycin treatment, PN(-/-) mice showed resistance to these changes. In vitro, dermal fibroblasts from PN(-/-) mice showed reduced transcript expression of alpha smooth actin and procollagen type-I alpha 1 (Col1α1) induced by transforming growth factor beta 1 (TGFβ1). Furthermore, recombinant mouse periostin directly induced Col1α1 expression in vitro, and this effect was inhibited by blocking the αv integrin-mediated PI3K/Akt signaling either with anti-αv functional blocking antibody or with the PI3K/Akt kinase inhibitor LY294002.

Conclusion: Periostin plays an essential role in the pathogenesis of Bleomycin-induced scleroderma in mice. Periostin may represent a potential therapeutic target for human scleroderma.

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Periostin upregulates the expression of Col1α1 via αv-integrin mediated-PI3K/Akt signaling pathway in vitro.A, Real-time quantitative PCR was performed to determine relative mRNA levels of Col1α1 in cultured dermal fibroblasts from WT and PN−/− mice after TGFβ1 stimulation at the indicated times. B, Relative mRNA levels of Col1α1 in WT mouse dermal fibroblasts with the indicated stimulation. C, Relative mRNA levels of Col1α1 in cultured WT mouse dermal fibroblasts treated with rmPeriostin in the presence or absence of the indicated neutralizing antibody or kinase inhibitors. D, Phosphorylation of Akt in cultured WT mouse dermal fibroblasts treated with or without rmPeriostin in the presence or absence of LY294002 or anti-αv neutralizing antibody. Values in A, B, and C were normalized to GAPDH levels and expressed as relative mRNA levels compared with WT mice fibroblasts (A) or WT dermal fibroblasts without stimulation (B and C). NS, no significance; ***, p<0.01.
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pone-0041994-g006: Periostin upregulates the expression of Col1α1 via αv-integrin mediated-PI3K/Akt signaling pathway in vitro.A, Real-time quantitative PCR was performed to determine relative mRNA levels of Col1α1 in cultured dermal fibroblasts from WT and PN−/− mice after TGFβ1 stimulation at the indicated times. B, Relative mRNA levels of Col1α1 in WT mouse dermal fibroblasts with the indicated stimulation. C, Relative mRNA levels of Col1α1 in cultured WT mouse dermal fibroblasts treated with rmPeriostin in the presence or absence of the indicated neutralizing antibody or kinase inhibitors. D, Phosphorylation of Akt in cultured WT mouse dermal fibroblasts treated with or without rmPeriostin in the presence or absence of LY294002 or anti-αv neutralizing antibody. Values in A, B, and C were normalized to GAPDH levels and expressed as relative mRNA levels compared with WT mice fibroblasts (A) or WT dermal fibroblasts without stimulation (B and C). NS, no significance; ***, p<0.01.

Mentions: TGFβ1 is also known as a major inducer of collagen synthesis. We therefore investigated Col1α1 transcript levels in WT and PN−/− fibroblasts when they were stimulated with TGFβ1. Similar to the results of α-SMA expression, Col1α1 expression in PN−/− fibroblasts became to be significantly lower than WT fibroblasts after 12 hours of stimulation (Figure 6A). This result suggests that periostin may play a role in the Col1α1 expression.


Periostin facilitates skin sclerosis via PI3K/Akt dependent mechanism in a mouse model of scleroderma.

Yang L, Serada S, Fujimoto M, Terao M, Kotobuki Y, Kitaba S, Matsui S, Kudo A, Naka T, Murota H, Katayama I - PLoS ONE (2012)

Periostin upregulates the expression of Col1α1 via αv-integrin mediated-PI3K/Akt signaling pathway in vitro.A, Real-time quantitative PCR was performed to determine relative mRNA levels of Col1α1 in cultured dermal fibroblasts from WT and PN−/− mice after TGFβ1 stimulation at the indicated times. B, Relative mRNA levels of Col1α1 in WT mouse dermal fibroblasts with the indicated stimulation. C, Relative mRNA levels of Col1α1 in cultured WT mouse dermal fibroblasts treated with rmPeriostin in the presence or absence of the indicated neutralizing antibody or kinase inhibitors. D, Phosphorylation of Akt in cultured WT mouse dermal fibroblasts treated with or without rmPeriostin in the presence or absence of LY294002 or anti-αv neutralizing antibody. Values in A, B, and C were normalized to GAPDH levels and expressed as relative mRNA levels compared with WT mice fibroblasts (A) or WT dermal fibroblasts without stimulation (B and C). NS, no significance; ***, p<0.01.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3404023&req=5

pone-0041994-g006: Periostin upregulates the expression of Col1α1 via αv-integrin mediated-PI3K/Akt signaling pathway in vitro.A, Real-time quantitative PCR was performed to determine relative mRNA levels of Col1α1 in cultured dermal fibroblasts from WT and PN−/− mice after TGFβ1 stimulation at the indicated times. B, Relative mRNA levels of Col1α1 in WT mouse dermal fibroblasts with the indicated stimulation. C, Relative mRNA levels of Col1α1 in cultured WT mouse dermal fibroblasts treated with rmPeriostin in the presence or absence of the indicated neutralizing antibody or kinase inhibitors. D, Phosphorylation of Akt in cultured WT mouse dermal fibroblasts treated with or without rmPeriostin in the presence or absence of LY294002 or anti-αv neutralizing antibody. Values in A, B, and C were normalized to GAPDH levels and expressed as relative mRNA levels compared with WT mice fibroblasts (A) or WT dermal fibroblasts without stimulation (B and C). NS, no significance; ***, p<0.01.
Mentions: TGFβ1 is also known as a major inducer of collagen synthesis. We therefore investigated Col1α1 transcript levels in WT and PN−/− fibroblasts when they were stimulated with TGFβ1. Similar to the results of α-SMA expression, Col1α1 expression in PN−/− fibroblasts became to be significantly lower than WT fibroblasts after 12 hours of stimulation (Figure 6A). This result suggests that periostin may play a role in the Col1α1 expression.

Bottom Line: Periostin, a novel matricellular protein, is recently reported to play a crucial role in tissue remodeling and is highly expressed under fibrotic conditions.To determine the downstream pathway involved in periostin signaling, receptor neutralizing antibody and signal transduction inhibitors were used in vitro.Furthermore, recombinant mouse periostin directly induced Col1α1 expression in vitro, and this effect was inhibited by blocking the αv integrin-mediated PI3K/Akt signaling either with anti-αv functional blocking antibody or with the PI3K/Akt kinase inhibitor LY294002.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Osaka University Graduate School of Medicine, Osaka, Japan.

ABSTRACT

Objective: Periostin, a novel matricellular protein, is recently reported to play a crucial role in tissue remodeling and is highly expressed under fibrotic conditions. This study was undertaken to assess the role of periostin in scleroderma.

Methods: Using skin from patients and healthy donors, the expression of periostin was assessed by immunohistochemistry and immunoblotting analyses. Furthermore, we investigated periostin(-/-) (PN(-/-)) and wild-type (WT) mice to elucidate the role of periostin in scleroderma. To induce murine cutaneous sclerosis, mice were subcutaneously injected with bleomycin, while untreated control groups were injected with phosphate-buffered saline. Bleomycin-induced fibrotic changes were compared in PN(-/-) and WT mice by histological analysis as well as by measurements of profibrotic cytokine and extracellular matrix protein expression levels in vivo and in vitro. To determine the downstream pathway involved in periostin signaling, receptor neutralizing antibody and signal transduction inhibitors were used in vitro.

Results: Elevated expression of periostin was observed in the lesional skin of patients with scleroderma compared with healthy donors. Although WT mice showed marked cutaneous sclerosis with increased expression of periostin and increased numbers of myofibroblasts after bleomycin treatment, PN(-/-) mice showed resistance to these changes. In vitro, dermal fibroblasts from PN(-/-) mice showed reduced transcript expression of alpha smooth actin and procollagen type-I alpha 1 (Col1α1) induced by transforming growth factor beta 1 (TGFβ1). Furthermore, recombinant mouse periostin directly induced Col1α1 expression in vitro, and this effect was inhibited by blocking the αv integrin-mediated PI3K/Akt signaling either with anti-αv functional blocking antibody or with the PI3K/Akt kinase inhibitor LY294002.

Conclusion: Periostin plays an essential role in the pathogenesis of Bleomycin-induced scleroderma in mice. Periostin may represent a potential therapeutic target for human scleroderma.

Show MeSH
Related in: MedlinePlus