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Scleroderma dermal fibroblasts overexpress vascular endothelial growth factor due to autocrine transforming growth factor β signaling

  • Original Article
  • Published:
Modern Rheumatology

Abstract

Objectives

Overexpression of vascular endothelial growth factor (VEGF) in scleroderma (SSc) skin may play a role in the pathogenesis of the disease. Our study was undertaken to evaluate whether dermal fibroblasts function as one of the sources of the increased VEGF in SSc, and to clarify its mechanism.

Methods

Protein and mRNA levels of VEGF were analyzed using immunoblotting, enzyme-linked immunosorbent assay, and real-time PCR. The DNA-binding ability of Smad3 was evaluated by DNA affinity precipitation.

Results

VEGF mRNA expression in vivo was increased in SSc skin compared to skin with other collagen diseases. Expression of VEGF protein and mRNA in cultured SSc dermal fibroblasts was constitutively and significantly upregulated. Ectopic TGF-β stimulation induced VEGF synthesis in normal fibroblasts, and TGF-β knockdown normalized the upregulated VEGF levels in SSc fibroblasts. Furthermore, Smad3 overexpression induced VEGF levels. We found that bp −532 to −521 on the VEGF promoter is a putative binding site for Smads, and that the binding activity of Smad3 to VEGF promoter was constitutively increased in SSc fibroblasts as well as in normal fibroblasts treated with exogenous TGF-β1.

Conclusions

We demonstrated that VEGF were overexpressed due to autocrine TGF-β/Smad signaling in SSc. TGF-β signaling may contribute to the pathogenesis of angiopathy as well as tissue fibrosis.

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Acknowledgments

We thank Mr. Keitaro Yamane, Ms. Junko Suzuki, and Ms. Chiemi Shiotsu for their valuable technical assistance. This study was supported in part by a Grant for Scientific Research from the Japanese Ministry of Education, Science, Sports and Culture, and by a research project on intractable diseases from the Japanese Ministry of Health, Labour and Welfare.

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None.

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Authors and Affiliations

  1. Department of Dermatology and Plastic Surgery, Faculty of Life Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, Japan

    Ikko Kajihara, Masatoshi Jinnin, Noritoshi Honda, Katsunari Makino, Takamitsu Makino, Shinichi Masuguchi, Keisuke Sakai, Satoshi Fukushima, Yuji Inoue & Hironobu Ihn

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  1. Ikko Kajihara
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  2. Masatoshi Jinnin
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  3. Noritoshi Honda
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  4. Katsunari Makino
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  5. Takamitsu Makino
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  8. Satoshi Fukushima
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  10. Hironobu Ihn
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Corresponding author

Correspondence to Masatoshi Jinnin.

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Kajihara, I., Jinnin, M., Honda, N. et al. Scleroderma dermal fibroblasts overexpress vascular endothelial growth factor due to autocrine transforming growth factor β signaling. Mod Rheumatol 23, 516–524 (2013). https://doi.org/10.1007/s10165-012-0698-6

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  • DOI: https://doi.org/10.1007/s10165-012-0698-6

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