Production of serum amyloid A in equine articular chondrocytes and fibroblast-like synoviocytes treated with proinflammatory cytokines and its effects on the two cell types in culture.
Abstract: To investigate the role of the major equine acute phase protein serum amyloid A (SAA) in inflammation of equine intraarticular tissues. Methods: Articular chondrocytes and fibroblast-like synoviocytes (FLSs) from 8 horses (4 horses/cell type). Methods: Chondrocytes and FLSs were stimulated in vitro for various periods up to 48 hours with cytokines (recombinant interleukin [IL]-1β, IL-6, tumor necrosis factor-α, or a combination of all 3 [IIT]) or with recombinant SAA. Gene expression of SAA, IL-6, matrix metalloproteinases (MMP)-1 and -3, and cartilage-derived retinoic acid-sensitive protein were assessed by quantitative real-time PCR assay; SAA protein was evaluated by immunoturbidimetry and denaturing isoelectric focusing and western blotting. Results: All cytokine stimulation protocols increased expression of SAA mRNA and resulted in detectable SAA protein production in chondrocytes and FLSs. Isoforms of SAA in lysed chondrocytes and their culture medium corresponded to those previously detected in synovial fluid from horses with joint disease. When exposed to SAA, chondrocytes and FLSs had increased expression of IL-6, SAA, and MMP3, and chondrocytes had increased expression of MMP-1. Chondrocytes had decreased expression of cartilage-derived retinoic acid-sensitive protein. Conclusions: Upregulation of SAA in chondrocytes and FLSs stimulated with proinflammatory cytokines and the proinflammatory effects of SAA suggested that SAA may be involved in key aspects of pathogenesis of the joint inflammation in horses.
Publication Date: 2015-12-29 PubMed ID: 26709936DOI: 10.2460/ajvr.77.1.50Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigates the role of serum amyloid A (SAA), a major acute phase protein in horses, and its involvement in inflammation of the horses’ intraarticular tissues. The study observed that increased levels of SAA are present in tissue cells (chondrocytes and fibroblast-like synoviocytes) when stimulated by proinflammatory cytokines and the presence of SAA increased the effects of inflammation.
Methods applied in the research
- The researchers used articular chondrocytes and fibroblast-like synoviocytes (FLSs) from 8 horses (4 each) for the study.
- The cells were stimulated in vitro for periods up to 48 hours using cytokines or with recombinant SAA.
- The expression of SAA, IL-6, matrix metalloproteinases (MMP)-1 and -3 was studied using a quantitative real-time PCR assay.
- Further, SAA protein was evaluated by immunoturbidimetry and denaturing isoelectric focusing and western blotting.
Observations and results
- All protocols involving cytokine stimulation saw an increase in the expression of SAA mRNA and SAA protein in both types of cells (chondrocytes and FLSs).
- The study found that the isoforms of SAA in chondrocytes and their culture medium are akin to those found in the synovial fluid of horses with joint diseases.
- Exposure to SAA was seen to increase the expression of IL-6, SAA, and MMP3 in chondrocytes and FLSs. Furthermore, in chondrocytes, there was an enhancement in the expression of MMP-1 and a reduction in the expression of cartilage-derived retinoic acid-sensitive protein.
Conclusions drawn from the study
- The researchers concluded that the upregulation of SAA in chondrocytes and FLSs, stimulated with proinflammatory cytokines, along with the proinflammatory effects of SAA itself, suggests that SAA might play a crucial role in the pathogenesis of joint inflammation in horses.
Cite This Article
APA
Jacobsen S, Ladefoged S, Berg LC.
(2015).
Production of serum amyloid A in equine articular chondrocytes and fibroblast-like synoviocytes treated with proinflammatory cytokines and its effects on the two cell types in culture.
Am J Vet Res, 77(1), 50-58.
https://doi.org/10.2460/ajvr.77.1.50 Publication
Researcher Affiliations
MeSH Terms
- Animals
- Cartilage / metabolism
- Cells, Cultured
- Chondrocytes / classification
- Chondrocytes / metabolism
- Cytokines / metabolism
- Cytokines / pharmacology
- Fibroblasts / drug effects
- Horses / genetics
- Horses / metabolism
- Interleukin-1beta / genetics
- Interleukin-6 / metabolism
- RNA, Messenger / metabolism
- Real-Time Polymerase Chain Reaction
- Serum Amyloid A Protein / metabolism
- Synovial Fluid / metabolism
- Synovial Membrane / cytology
- Synovial Membrane / metabolism
- Tumor Necrosis Factor-alpha / metabolism
Citations
This article has been cited 7 times.- Witkowska-Piłaszewicz OD, Żmigrodzka M, Winnicka A, Miśkiewicz A, Strzelec K, Cywińska A. Serum amyloid A in equine health and disease. Equine Vet J 2019 May;51(3):293-298.
- Colbath AC, Dow SW, Hopkins LS, Phillips JN, McIlwraith CW, Goodrich LR. Induction of Synovitis Using Interleukin-1 Beta: Are There Differences in the Response of Middle Carpal Joint Compared to the Tibiotarsal Joint?. Front Vet Sci 2018;5:208.
- Ghasemi S, Sardari K, Mirshokraei P, Hassanpour H. In vitro study of matrix metalloproteinases 1, 2, 9, 13 and serum amyloid A mRNAs expression in equine fibroblast-like synoviocytes treated with doxycycline. Can J Vet Res 2018 Apr;82(2):82-88.
- Andreassen SM, Vinther AML, Nielsen SS, Andersen PH, Tnibar A, Kristensen AT, Jacobsen S. Changes in concentrations of haemostatic and inflammatory biomarkers in synovial fluid after intra-articular injection of lipopolysaccharide in horses. BMC Vet Res 2017 Jun 19;13(1):182.
- Lu T, Liu ZY, Ge YS, Jiang SY, Zhao QA, Ding DF. Study of Retinoic Acid-Induced Osteoarthritis: Integrating RNA-Sequencing, Network Pharmacology, Molecular Docking, and Experimental Validation. Int J Mol Sci 2025 Jun 9;26(12).
- Dayı B, Sevimli A, Akkoç A, Mutlu AM, Doğan N. Serum amyloid a production in chicken embryonic synovial fibroblasts induced by lipopolysaccharide, interleukin-1β, and vitamin A. Poult Sci 2025 Sep;104(9):105358.
- Koziy RV, Katselis GS, Yoshimura S, Simko E, Bracamonte JL. Temporal kinetics of serum amyloid A (SAA) concentration and identification of SAA isoforms in blood and synovial fluid of horses with experimentally induced septic arthritis, non-septic synovitis, and systemic inflammation. J Vet Diagn Invest 2025 Jan;37(1):42-54.
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