Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes.
Abstract: To determine the effects of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) on expression and regulation of several matrix-related genes by equine articular chondrocytes. Methods: Articular cartilage harvested from grossly normal joints of 8 foals, 6 yearling horses, and 8 adult horses. Methods: Chondrocytes maintained in suspension cultures were treated with various doses of human recombinant IL-1beta or TNF-alpha. Northern blots of total RNA from untreated and treated chondrocytes were probed with equine complementary DNA (cDNA) probes for cartilage matrix-related genes. Incorporation of 35S-sulfate, fluorography of 14C-proline labeled medium, zymography, and western blotting were used to confirm effects on protein synthesis. Results: IL-1beta and TNF-alpha increased steady-state amounts of mRNA of matrix metalloproteinases 1, 3, and 13 by up to 100-fold. Amount of mRNA of tissue inhibitor of metalloproteinase-1 also increased but to a lesser extent (1.5- to 2-fold). Amounts of mRNA of type-II collagen and link protein were consistently decreased in a dose-dependent manner. Amount of aggrecan mRNA was decreased slightly; amounts of biglycan and decorin mRNA were minimally affected. Conclusions: Treatment of cultured equine chondrocytes with IL-1beta or TNF-alpha resulted in marked alterations in expression of various matrix and matrix-related genes consistent with the implicated involvement of these genes in arthritis. Expression of matrix metalloproteinases was increased far more than expression of their putative endogenous inhibitor. Results support the suggestion that IL-1beta and TNF-alpha play a role in the degradation of articular cartilage in arthritis.
Publication Date: 2000-06-13 PubMed ID: 10850836DOI: 10.2460/ajvr.2000.61.624Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study explored how interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha), two biochemical agents, influence the expression of certain matrix-related genes in horse cartilage cells. The results showed that these agents considerably alter gene expression and could be involved in the progression of arthritis, as they potentially contribute to the degradation of joint cartilage.
Study Design and Methodology
The researchers conducted the experimental study on articular cartilage taken from healthy joints of eight foals, six yearling horses, and eight adult horses. Specifically, they:
- Kept the chondrocytes (cartilage cells) in suspension cultures and treated them with varying doses of the biochemical agents IL-1beta or TNF-alpha.
- Used Northern blots of the RNA taken from both untreated and treated chondrocytes, probing them with horse DNA sequences for cartilage matrix-related genes.
- Verified the effects on protein synthesis using 35S-sulfate incorporation, fluorography of 14C-proline labeled medium, zymography, and western blotting techniques.
Key Findings
The results indicated that:
- IL-1beta and TNF-alpha increased steady-state quantities of mRNA of matrix metalloproteinases (enzymes that break down proteins) 1, 3, and 13 by up to 100 times.
- The mRNA amount of the tissue inhibitor of metalloproteinase-1, an agent that inhibits matrix metalloproteinases, increased as well, but only to a lesser extent (1.5- to 2-fold).
- The mRNA amounts of type-II collagen and link protein consistently decreased in a dose-dependent manner.
- The amount of aggrecan mRNA slightly declined, whereas the amounts of biglycan and decorin mRNA were minimally affected.
Conclusions and Implications
The treatment of horse chondrocytes with IL-1beta or TNF-alpha led to substantial alterations in the expression of various matrix and matrix-related genes. This aligns with the suspected involvement of these genes in arthritis progression, as the increase in matrix metalloproteinases was far higher than their supposed endogenous inhibitor. This suggests that IL-1beta and TNF-alpha potentially contribute to the degradation of articular cartilage in arthritis. These findings support the possibility of these agents being potential targets for therapeutic interventions in arthritis.
Cite This Article
APA
Richardson DW, Dodge GR.
(2000).
Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes.
Am J Vet Res, 61(6), 624-630.
https://doi.org/10.2460/ajvr.2000.61.624 Publication
Researcher Affiliations
- Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square 19348-1692, USA.
MeSH Terms
- Animals
- Blotting, Northern / veterinary
- Blotting, Western / veterinary
- Cartilage, Articular / chemistry
- Cartilage, Articular / cytology
- Cartilage, Articular / physiology
- Chondrocytes / chemistry
- Chondrocytes / physiology
- Collagenases / genetics
- Collagenases / physiology
- Electrophoresis, Polyacrylamide Gel / veterinary
- Fluorometry / veterinary
- Gene Expression Regulation, Enzymologic
- Glycosaminoglycans / analysis
- Horses / genetics
- Horses / physiology
- Image Processing, Computer-Assisted
- Interleukin-1 / genetics
- Interleukin-1 / physiology
- Matrix Metalloproteinase 1 / genetics
- Matrix Metalloproteinase 1 / physiology
- Matrix Metalloproteinase 13
- Matrix Metalloproteinase 3 / genetics
- Matrix Metalloproteinase 3 / physiology
- Matrix Metalloproteinases / genetics
- Matrix Metalloproteinases / metabolism
- Scintillation Counting
- Statistics, Nonparametric
- Sulfates / chemistry
- Tumor Necrosis Factor-alpha / genetics
- Tumor Necrosis Factor-alpha / physiology
Grant Funding
- AR42417 / NIAMS NIH HHS
Citations
This article has been cited 32 times.- Selig M, Azizi S, Walz K, Lauer JC, Rolauffs B, Hart ML. Cell morphology as a biological fingerprint of chondrocyte phenotype in control and inflammatory conditions. Front Immunol 2023;14:1102912.
- Harvanova D, Matejova J, Slovinska L, Lacko M, Gulova S, Fecskeova LK, Janockova J, Spakova T, Rosocha J. The Role of Synovial Membrane in the Development of a Potential In Vitro Model of Osteoarthritis. Int J Mol Sci 2022 Feb 24;23(5).
- Hotham WE, Thompson C, Szu-Ting L, Henson FMD. The anti-inflammatory effects of equine bone marrow stem cell-derived extracellular vesicles on autologous chondrocytes. Vet Rec Open 2021 Dec;8(1):e22.
- Wen ZQ, Liu D, Zhang Y, Cai ZJ, Xiao WF, Li YS. G Protein-Coupled Receptors in Osteoarthritis: A Novel Perspective on Pathogenesis and Treatment. Front Cell Dev Biol 2021;9:758220.
- Bartolotti I, Roseti L, Petretta M, Grigolo B, Desando G. A Roadmap of In Vitro Models in Osteoarthritis: A Focus on Their Biological Relevance in Regenerative Medicine. J Clin Med 2021 Apr 28;10(9).
- Jayakumar T, Saravana Bhavan P, Sheu JR. Molecular Targets of Natural Products for Chondroprotection in Destructive Joint Diseases. Int J Mol Sci 2020 Jul 13;21(14).
- Sirikaew N, Chomdej S, Tangyuenyong S, Tangjitjaroen W, Somgird C, Thitaram C, Ongchai S. Proinflammatory cytokines and lipopolysaccharides up regulate MMP-3 and MMP-13 production in Asian elephant (Elephas maximus) chondrocytes: attenuation by anti-arthritic agents. BMC Vet Res 2019 Nov 21;15(1):419.
- Kardos D, Marschall B, Simon M, Hornyák I, Hinsenkamp A, Kuten O, Gyevnár Z, Erdélyi G, Bárdos T, Paukovits TM, Magos K, Béres G, Szenthe K, Bánáti F, Szathmary S, Nehrer S, Lacza Z. Investigation of Cytokine Changes in Osteoarthritic Knee Joint Tissues in Response to Hyperacute Serum Treatment. Cells 2019 Aug 3;8(8).
- Cho IA, Kim TH, Lim H, Park JH, Kang KR, Lee SY, Kim CS, Kim DK, Kim HJ, Yu SK, Kim SG, Kim JS. Formononetin Antagonizes the Interleukin-1β-Induced Catabolic Effects Through Suppressing Inflammation in Primary Rat Chondrocytes. Inflammation 2019 Aug;42(4):1426-1440.
- Kim TH, Yun YP, Shim KS, Kim HJ, Kim SE, Park K, Song HR. In Vitro Anti-Inflammation and Chondrogenic Differentiation Effects of Inclusion Nanocomplexes of Hyaluronic Acid-Beta Cyclodextrin and Simvastatin. Tissue Eng Regen Med 2018 Jun;15(3):263-274.
- Castro Martins M, Peffers MJ, Lee K, Rubio-Martinez LM. Effects of stanozolol on normal and IL-1β-stimulated equine chondrocytes in vitro. BMC Vet Res 2018 Mar 20;14(1):103.
- Pan L, Zhang Y, Chen N, Yang L. Icariin Regulates Cellular Functions and Gene Expression of Osteoarthritis Patient-Derived Human Fibroblast-Like Synoviocytes. Int J Mol Sci 2017 Dec 8;18(12).
- Byron CR, Trahan RA. Comparison of the Effects of Interleukin-1 on Equine Articular Cartilage Explants and Cocultures of Osteochondral and Synovial Explants. Front Vet Sci 2017;4:152.
- Wan Q, TruongVo T, Steele HE, Ozcelikkale A, Han B, Wang Y, Oh J, Yokota H, Na S. Subcellular domain-dependent molecular hierarchy of SFK and FAK in mechanotransduction and cytokine signaling. Sci Rep 2017 Aug 22;7(1):9033.
- Lee AS, Ellman MB, Yan D, Kroin JS, Cole BJ, van Wijnen AJ, Im HJ. A current review of molecular mechanisms regarding osteoarthritis and pain. Gene 2013 Sep 25;527(2):440-7.
- Murab S, Chameettachal S, Bhattacharjee M, Das S, Kaplan DL, Ghosh S. Matrix-embedded cytokines to simulate osteoarthritis-like cartilage microenvironments. Tissue Eng Part A 2013 Aug;19(15-16):1733-53.
- Scanzello CR, Goldring SR. The role of synovitis in osteoarthritis pathogenesis. Bone 2012 Aug;51(2):249-57.
- Sun L, Wang X, Kaplan DL. A 3D cartilage - inflammatory cell culture system for the modeling of human osteoarthritis. Biomaterials 2011 Aug;32(24):5581-9.
- Umlauf D, Frank S, Pap T, Bertrand J. Cartilage biology, pathology, and repair. Cell Mol Life Sci 2010 Dec;67(24):4197-211.
- Millward-Sadler SJ, Costello PW, Freemont AJ, Hoyland JA. Regulation of catabolic gene expression in normal and degenerate human intervertebral disc cells: implications for the pathogenesis of intervertebral disc degeneration. Arthritis Res Ther 2009;11(3):R65.
- Fan Z, Söder S, Oehler S, Fundel K, Aigner T. Activation of interleukin-1 signaling cascades in normal and osteoarthritic articular cartilage. Am J Pathol 2007 Sep;171(3):938-46.
- Fan Z, Yang H, Bau B, Söder S, Aigner T. Role of mitogen-activated protein kinases and NFkappaB on IL-1beta-induced effects on collagen type II, MMP-1 and 13 mRNA expression in normal articular human chondrocytes. Rheumatol Int 2006 Aug;26(10):900-3.
- Le Maitre CL, Freemont AJ, Hoyland JA. The role of interleukin-1 in the pathogenesis of human intervertebral disc degeneration. Arthritis Res Ther 2005;7(4):R732-45.
- Wheaton AJ, Dodge GR, Borthakur A, Kneeland JB, Schumacher HR, Reddy R. Detection of changes in articular cartilage proteoglycan by T(1rho) magnetic resonance imaging. J Orthop Res 2005 Jan;23(1):102-8.
- Tung JT, Fenton JI, Arnold C, Alexander L, Yuzbasiyan-Gurkan V, Venta PJ, Peters TL, Orth MW, Richardson DW, Caron JP. Recombinant equine interleukin-1beta induces putative mediators of articular cartilage degradation in equine chondrocytes. Can J Vet Res 2002 Jan;66(1):19-25.
- Long P, Gassner R, Agarwal S. Tumor necrosis factor alpha-dependent proinflammatory gene induction is inhibited by cyclic tensile strain in articular chondrocytes in vitro. Arthritis Rheum 2001 Oct;44(10):2311-9.
- Smith KWY, Fung SL, Wu HF, Chiesa I, Vozzi G, De Maria C, Gottardi R. Developing an in vitro osteochondral micro-physiological system for modeling cartilage-bone crosstalk in arthritis. Front Immunol 2025;16:1495613.
- Zhang M, Xiong W, Qiao R, Li M, Zhang C, Yang C, Zhu Y, He J, Ma Z. Irisin in the modulation of bone and cartilage homeostasis: a review on osteoarthritis relief potential. Front Physiol 2025;16:1570157.
- Teo KYW, Sevencan C, Cheow YA, Zhang S, Leong DT, Toh WS. Macrophage Polarization as a Facile Strategy to Enhance Efficacy of Macrophage Membrane-Coated Nanoparticles in Osteoarthritis. Small Sci 2022 Apr;2(4):2100116.
- Imagawa M, Shinjo T, Sato K, Kawakami K, Zeze T, Nishimura Y, Toyoda M, Chen S, Ryo N, Ahmed AK, Iwashita M, Yamashita A, Fukuda T, Sanui T, Nishimura F. Epithelial-to-mesenchymal transition, inflammation, subsequent collagen production, and reduced proteinase expression cooperatively contribute to cyclosporin-A-induced gingival overgrowth development. Front Physiol 2023;14:1298813.
- Liu B, Wu Y, Liang T, Zhou Y, Chen G, He J, Ji C, Liu P, Zhang C, Lin J, Shi K, Luo Z, Liu N, Su X. Betulinic Acid Attenuates Osteoarthritis via Limiting NLRP3 Inflammasome Activation to Decrease Interleukin-1β Maturation and Secretion. Mediators Inflamm 2023;2023:3706421.
- Chen L, Yu Y. Exercise and Osteoarthritis. Adv Exp Med Biol 2020;1228:219-231.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
