Anti-fibrotic Actions of Equine Interleukin-10 on Transforming Growth Factor-Beta1-Stimulated Dermal Fibroblasts Isolated From Limbs of Horses.
Abstract: Fibroproliferative disorders occur in both humans and horses following skin injury. In horses, wound healing on the limb is often complicated by the formation of fibroproliferative exuberant granulation tissue, characterized by persistent expression of pro-fibrotic transforming growth factor-beta1 (TGF-β1) and deficient expression of anti-inflammatory interleukin-10 (IL-10). IL-10 has been shown to directly modulate fibrotic gene expression in human fibroblasts, so we hypothesized that equine IL-10 (eIL-10) may exert similar anti-fibrotic effects on equine dermal fibroblasts. Cell-lines were created from the limb skin of six individual horses. Recombinant eIL-10 was produced and purified, and its effects on the cells investigated in the presence and absence of equine TGF-β1 (eTGF-β1). Myofibroblast differentiation and collagen production were examined using immunofluorescent cytometry, cell contractility in a collagen gel assay, and fibrotic gene expression using quantitative PCR. In response to eTGF-β1, fibroblasts increased in contractility and expression of alpha-smooth muscle actin, collagen types 1 and 3, and matrix metalloproteinase 1, 2, and 9. Equine IL-10 limited cell contractility and production of alpha-smooth muscle actin and type 3 collagen, and decreased mRNA levels of eCol3a1 and eMMP9, while increasing that of eMMP1. Opposing effects on eTGF-βR3 and eIL-10R1 gene expression were also observed, with mRNA levels decreasing following eTGF-β1 treatment, and increasing with eIL-10 treatment. These findings indicate that eIL-10 limits the pro-fibrotic effects of eTGF-β1, potentially through the modulation of fibrotic and receptor gene expression. Further investigations are warranted to assess the therapeutic utility of eIL-10 in the treatment of exuberant granulation tissue.
Copyright © 2020 Wise, Stuart, Sriutaisuk, Adams, Riley and Theoret.
Publication Date: 2020-09-18 PubMed ID: 33195583PubMed Central: PMC7531226DOI: 10.3389/fvets.2020.577835Google Scholar: Lookup
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Summary
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The research article explores the anti-fibrotic effects of equine interleukin-10 (eIL-10) on fibroblast cells from horse skin. The study supports the idea that eIL-10 can counteract the fibrotic influence of transforming growth factor-beta1 (TGF-β1), suggesting potential therapeutic applications.
Why the Research?
- This research was conducted in response to an observed problem in horses: wound healing on the limbs often results in the formation of fibroproliferative exuberant granulation tissues, which is a disorder associated with excessive scar tissue.
- The continuous expression of pro-fibrotic transforming growth factor-beta1 (TGF-β1) and insufficient expression of anti-inflammatory interleukin-10 (IL-10) are defining characteristics of these fibroproliferative disorders.
- Previous research showed that IL-10 can alter fibrotic gene expression in human fibroblasts, which leads to the hypothesis that equine IL-10 (eIL-10) might have a similar effect on equine dermal fibroblasts.
The Experiment
- Fibroblasts were derived from the skin of six different horses to create cell lines. Recombinant eIL-10 was prepared and assessed for its impact on these cells, both with and without the presence of equine TGF-β1 (eTGF-β1).
- The researchers examined myofibroblast differentiation, collagen production, and fibrotic gene expression, employing techniques such as immunofluorescent cytometry, a collagen gel assay for cell contractility, and quantitative PCR for gene expression analysis.
Findings
- Fibroblasts showed an increase in contractility and the expression of alpha-smooth muscle actin, collagen types 1 and 3, and matrix metalloproteinase 1, 2, and 9 in response to eTGF-β1.
- However, eIL-10 appeared to curb cell contractility and the production of alpha-smooth muscle actin and type 3 collagen. It also decreased mRNA levels of certain fibrotic genes while increasing those of others.
- It was suggested, therefore, that eIL-10 can limit the pro-fibrotic effects of eTGF-β1, possibly through the modulation of fibrotic and receptor gene expression.
What Next?
- These encouraging findings warrant further investigation to ascertain the potential therapeutic applications of eIL-10 in treating horses’ exuberant granulation tissue, and perhaps, by extension, similar conditions in humans.
Cite This Article
APA
Wise LM, Stuart GS, Sriutaisuk K, Adams BR, Riley CB, Theoret CL.
(2020).
Anti-fibrotic Actions of Equine Interleukin-10 on Transforming Growth Factor-Beta1-Stimulated Dermal Fibroblasts Isolated From Limbs of Horses.
Front Vet Sci, 7, 577835.
https://doi.org/10.3389/fvets.2020.577835 Publication
Researcher Affiliations
- Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand.
- Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand.
- Department of Pharmacology and Toxicology, University of Otago, Dunedin, New Zealand.
- School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- School of Veterinary Science, Massey University, Palmerston North, New Zealand.
- Département de Biomedecine Vétérinaire, Faculté de Médecine Vétérinaire, Université de Montréal, Montreal, QC, Canada.
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Citations
This article has been cited 1 times.- Naqash A, Stuart G, Kemp R, Wise L. Parapoxvirus Interleukin-10 Homologues Vary in Their Receptor Binding, Anti-Inflammatory, and Stimulatory Activities. Pathogens 2022 Apr 24;11(5).
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