Distribution of TNF receptors and TNF receptor-associated intracellular signaling factors on equine tendinocytes in vitro.
Abstract: Although tumor necrosis factor (TNF) alpha is an important key factor in degeneration of equine superficial digital flexor tendon (SDFT), the dynamism of TNF receptors and associated factors on tendinocytes has not been elucidated. To reveal signaling events mediated by TNF-receptors (TNF-Rs) in tendinocytes, we focused on four signaling factors, TNF-R1, TNF-R2, TNF-R-associated factor 2 (TRAF2) and nuclear factor-kappa B (NF-kappaB), and investigated the distribution and production of these factors. Cultured tendinocytes were obtained from SDFTs of thoroughbred horses. The tendinocytes were treated with 10 ng/ml equine TNFalpha medium for 6 hours and then the four factors on tendinocytes were visualized by using an immunohistochemical method, and the amounts of the four factors were determined by Western blot analysis. Although TNF-R1 and TNF-R2 co-localized on the same tendinocyte, in untreated control cells (normal condition), immunoreactivity against TNF -R1 was very weak but TNF-R2 showed a strong reaction. However, TNF-R1 showed the same high level of reaction as TNF-R2 in TNFalpha-treated cells (inflamed condition). Intense TRAF2 and NF-kappaB were detected at inflamed condition, however both factors were also detected at normal condition. The distinct distributions of the four factors under different conditions (normal and inflamed condition) in vitro not only reflect the dynamism of the cytokines but may also provide important clues for a means to prevent from occurrence of tendonitis and progress of tendon degeneration.
Publication Date: 2005-01-06 PubMed ID: 15631011
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research explores the distribution and behavior of certain tumor necrosis factor (TNF) receptors on equine tendinocytes, providing insight into tendon degeneration processes in horses.
Objective of the Research
- The main goal of this study was to investigate the dynamics of TNF receptors and their associated factors on tendinocytes. This is key to understanding the degeneration of the horse’s superficial digital flexor tendon (SDFT).
The Methodology
- Cultured tendinocytes were taken from the SDFT of thoroughbred horses.
- The tendinocytes were treated with an equine TNF alpha compound.
- Immunohistochemical methods were used to observe and study the presence and movement of four signaling factors. These factors included TNF-R1, TNF-R2, TNF-R-associated factor 2 (TRAF2), and nuclear factor-kappa B (NF-kappaB).
- The quantities of these factors were then established using Western blot analysis.
Key Findings
- While TNF-R1 and TNF-R2 were found to be present on the same tendinocyte, the reaction level of TNF-R1 was weak, while that of TNF-R2 was significantly stronger.
- When treated with TNFalpha, both receptors showed similar, high-levels of reactivity. This reactivity was observed in the “inflamed condition” of the cells.
- TRAF2 and NF-kappaB showed high levels of detection in inflamed cells, although they were also detected in normal conditions.
Significance of the Research
- This study offers a clearer understanding of how proteins and factors interact in tendon cells, particularly under normal and inflamed conditions.
- The findings therefore provide an insight into the mechanisms of tendon degeneration and could lead to innovative methods for preventing tendonitis and tendon degeneration.
Cite This Article
APA
Hosaka Y, Sakamoto Y, Kirisawa R, Watanabe T, Ueda H, Takehana K, Yamaguchi M.
(2005).
Distribution of TNF receptors and TNF receptor-associated intracellular signaling factors on equine tendinocytes in vitro.
Jpn J Vet Res, 52(3), 135-144.
Publication
Researcher Affiliations
- Department of Veterinary Anatomy, School of Veterinary Medicine, Rakuno Gakuen University, 582 Bunkyodai-Midorimachi, Ebetsu, Hokkaido 069-8501, Japan. hosap@rakuno.ac.jp
MeSH Terms
- Animals
- Blotting, Western / veterinary
- Cells, Cultured
- Female
- Horses / metabolism
- Immunohistochemistry / veterinary
- Male
- Microscopy, Fluorescence / veterinary
- NF-kappa B / metabolism
- Receptors, Tumor Necrosis Factor, Type I / metabolism
- Receptors, Tumor Necrosis Factor, Type II / metabolism
- TNF Receptor-Associated Factor 2 / metabolism
- Tendons / cytology
- Tendons / metabolism
Citations
This article has been cited 3 times.- Russo V, Mauro A, Martelli A, Di Giacinto O, Di Marcantonio L, Nardinocchi D, Berardinelli P, Barboni B. Cellular and molecular maturation in fetal and adult ovine calcaneal tendons. J Anat 2015 Feb;226(2):126-42.
- de Girolamo L, Viganò M, Galliera E, Stanco D, Setti S, Marazzi MG, Thiebat G, Corsi Romanelli MM, Sansone V. In vitro functional response of human tendon cells to different dosages of low-frequency pulsed electromagnetic field. Knee Surg Sports Traumatol Arthrosc 2015 Nov;23(11):3443-53.
- Lai C, Li R, Tang W, Liu J, Duan XDXF, Bao D, Liu H, Fu S. Metabolic Syndrome and Tendon Disease: A Comprehensive Review. Diabetes Metab Syndr Obes 2024;17:1597-1609.
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