Molecular and Cellular Biochemistry

, Volume 452, Issue 1–2, pp 177–185 | Cite as

Wound fluid of rats fed protein-free diets delays wound healing through the suppression of the IGF-1/ERK(1/2) signaling pathway

  • Takumi Yamane
  • Mari Shimura
  • Ryosuke Konno
  • Ken Iwatsuki
  • Yuichi OishiEmail author


Adequate nutrition is required to maintain healthy skin integrity, and malnourished patients with poor protein diet often experience delayed wound healing. Understanding the cellular mechanisms of protein malnutrition will justify the importance of optimal protein diets in health and disease defense. Therefore in the present study, we examined the effects of changes in wound fluid composition and its function caused by protein malnutrition on wound healing. Rats were fed a control (CO; 20% protein) diet or a protein-free (PF) diet for 2 weeks; we then created full-thickness wounds on the dorsolateral skin. On day 5 after wounding, frozen sections of the wounds were created to investigate the state of granulation tissues, and wound fluid obtained from the rats was collected to examine variations in cytokine levels and its function. Wound closure was significantly delayed from day 4 until total wound closure in rats fed a PF diet. Thickness of granulation tissue, which is composed of mainly dermal fibroblasts, and Ki67 immunohistochemical staining were significantly decreased in rats fed PF diets. PF diets decreased insulin-like growth factor (IGF)-I, which promotes wound healing, and increased IGF-binding protein-1, which inhibits IGF-I bioavailability, in wound fluid. Wound fluid obtained from rats fed a PF diet suppressed dermal fibroblast proliferation. Furthermore, the wound fluid remarkably decreased the phosphorylation level of IGF-I receptor β (IGF-IR) and extracellular signal-regulated kinase (ERK)(1/2) in dermal fibroblasts. These results show that wound fluid of rats fed PF diets delays wound healing by inhibiting granulation tissue formation through the suppression of the IGF-1/ERK(1/2) signaling pathway.


Dermal fibroblast Insulin-like growth factor-I Protein malnutrition Wound fluid Wound healing 



Extracellular signal-regulated kinase


Insulin-like growth factor


Hepatocyte growth factor




Macrophage colony-stimulating factor


Preadipocyte factor-1


Tumor necrosis factor-alpha


Monocyte chemoattractant protein-1


Vascular endothelial growth factor


Intercellular adhesion molecule-1


Dipeptidyl peptidase-4


IGF-binding protein


Receptor for advanced glycation endproducts


Leukemia inhibitory factor


Regulated on activation, normal T cell expressed and secreted


Tissue inhibitor of metalloproteinase 1


Fibroblast growth factor 21


Fetal bovine serum


Modified Eagle’s medium




Phosphate-buffered saline


Tris buffered saline containing 0.5% (v/v) Tween 20



We thank all of the persons who cooperated in this study. This work was supported by JSPS KAKENHI Grant Number 26861876.

Author contributions

MS and RK performed the research and analyzed the data; TY and MS wrote the paper; TY, KI, and YO discussed the data; TY designed the research study.

Compliance with ethical standards

Conflict of interest

The authors report no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Takumi Yamane
    • 1
  • Mari Shimura
    • 1
  • Ryosuke Konno
    • 1
  • Ken Iwatsuki
    • 1
  • Yuichi Oishi
    • 1
    Email author
  1. 1.Department of Nutritional Science and Food Safety, Faculty of Applied BioscienceTokyo University of AgricultureTokyoJapan

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