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Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2002; 66(1); 19-25;

Recombinant equine interleukin-1beta induces putative mediators of articular cartilage degradation in equine chondrocytes.

Abstract: Interleukin-1 is considered a central mediator of cartilage loss in osteoarthritis in several species, however an equine recombinant form of this cytokine is not readily available for in vitro use in equine osteoarthritis research. Equine recombinant interleukin-1beta was cloned and expressed and its effects on the expression and activity of selected chondrocytic proteins implicated in cartilage matrix degradation were characterized. Reverse transcriptase polymerase chain reaction methods were used to amplify the entire coding region of the equine IL-1beta mRNA, which was cloned into an expression vector, expressed in E. coli, and purified using a Ni2+ chromatographic method. The effects of the recombinant peptide on chondrocyte gene expression were determined by Northern blotting using RNA from equine chondrocyte cultures hybridized to probes for matrix metalloproteinases (MMP 1, MMP 3, MMP 13), tissue inhibitor of matrix metalloproteinases 1 (TIMP 1) and cyclooxygenase 2 (COX 2). Effects on selected mediators of cartilage degradation (nitrite concentrations and MMP activity) were determined using conditioned medium from reIL-1beta-treated equine cartilage explant cultures. A recombinant peptide of approximately 21 kd was obtained. Northern blotting analyses revealed a marked up-regulation of expression of all MMPs, TIMP 1, and COX 2 in mRNA from treated chondrocytes. Furthermore, cartilage explants exposed to reIL-1beta had augmented collagenase/gelatinase and stromelysin activities as well as increased concentration of nitrite in conditioned media. The development of a biologically active, species-specific IL-1beta provides a valuable tool in the study of osteoarthritis pathophysiology and its treatment in horses.
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Publication Date: 2002-02-23 PubMed ID: 11858644PubMed Central: PMC226977
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • Non-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 research is about the development and testing of a species-specific version of a protein called interleukin-1beta, which is believed to play a critical role in cartilage loss in osteoarthritis, specifically in horses.

Objective of the Research

The goal of this study was to produce and evaluate the effects of a horse-specific form of interleukin-1beta, a protein thought to be a key player in the degradation of cartilage in osteoarthritis. Since an equine version of this protein wasn’t readily available for lab work, the researchers sought to clone and express it themselves.

Methodology

  • The researchers used reverse transcriptase polymerase chain reaction methods to amplify the genetic coding for equine interleukin-1beta, which they then cloned into an expression vector.
  • They expressed the gene in E.coli and then purified the produced protein using a Ni2+ chromatographic method.
  • Once they had the protein, they evaluated its effects on the gene expression of horse chondrocytes (cartilage cells) by testing it on equine chondrocyte cultures and conducting Northern blotting to analyze the expression of matrix metalloproteinases (MMP 1, MMP 3, MMP 13) and other related genes.

Findings

  • The team successfully produced a recombinant peptide of about 21 kilodaltons in size.
  • Their tests revealed that treating the chondrocytes with the peptide resulted in a sharp increase in the expression of all tested matrix metalloproteinases and other associated genes. Matrix metalloproteinases are enzymes that break down the extracellular matrix in normal physiological processes.
  • Exposure to the peptide also boosted collagenase/gelatinase activities and elevated the concentration of nitrite in the conditioned media, indicating enhanced cartilage degradation.

Conclusion

The research team successfully developed an equine-specific version of interleukin-1beta and showed that it promotes the activity of factors implicated in cartilage loss. This form of the protein could serve as a vital tool in studying the pathophysiology of osteoarthritis in horses and in seeking potential treatments.

Cite This Article

APA
Tung JT, Fenton JI, Arnold C, Alexander L, Yuzbasiyan-Gurkan V, Venta PJ, Peters TL, Orth MW, Richardson DW, Caron JP. (2002). Recombinant equine interleukin-1beta induces putative mediators of articular cartilage degradation in equine chondrocytes. Can J Vet Res, 66(1), 19-25.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 0830-9000
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 66
Issue: 1
Pages: 19-25

Researcher Affiliations

Tung, J T
  • Department of Animal Science, Michigan State University, East Lansing 48824, USA.
Fenton, J I
    Arnold, C
      Alexander, L
        Yuzbasiyan-Gurkan, V
          Venta, P J
            Peters, T L
              Orth, M W
                Richardson, D W
                  Caron, J P

                    MeSH Terms

                    • Animals
                    • Blotting, Northern / veterinary
                    • Cartilage, Articular / drug effects
                    • Cartilage, Articular / enzymology
                    • Cartilage, Articular / pathology
                    • Cells, Cultured
                    • Chondrocytes / drug effects
                    • Chondrocytes / enzymology
                    • Cyclooxygenase 2
                    • Gene Expression Regulation, Enzymologic
                    • Horse Diseases / physiopathology
                    • Horses
                    • Interleukin-1 / pharmacology
                    • Interleukin-1 / physiology
                    • Isoenzymes / genetics
                    • Isoenzymes / metabolism
                    • Matrix Metalloproteinases / genetics
                    • Matrix Metalloproteinases / metabolism
                    • Osteoarthritis / physiopathology
                    • Osteoarthritis / veterinary
                    • Prostaglandin-Endoperoxide Synthases / genetics
                    • Prostaglandin-Endoperoxide Synthases / metabolism
                    • RNA, Messenger / metabolism
                    • Recombinant Proteins / metabolism
                    • Recombinant Proteins / pharmacology
                    • Reverse Transcriptase Polymerase Chain Reaction / veterinary
                    • Tissue Inhibitor of Metalloproteinase-1 / genetics
                    • Tissue Inhibitor of Metalloproteinase-1 / metabolism

                    References

                    This article includes 47 references
                    1. Martel-Pelletier J, Alaaeddine N, Pelletier JP. Cytokines and their role in the pathophysiology of osteoarthritis.. Front Biosci 1999 Oct 15;4:D694-703.
                      pubmed: 10525480doi: 10.2741/martelgoogle scholar: lookup
                    2. van den Berg WB. The role of cytokines and growth factors in cartilage destruction in osteoarthritis and rheumatoid arthritis.. Z Rheumatol 1999 Jun;58(3):136-41.
                      pubmed: 10441840doi: 10.1007/s003930050163google scholar: lookup
                    3. Pelletier JP, DiBattista JA, Roughley P, McCollum R, Martel-Pelletier J. Cytokines and inflammation in cartilage degradation.. Rheum Dis Clin North Am 1993 Aug;19(3):545-68.
                      pubmed: 8210574
                    4. Okada Y. Proteinases and matrix degradation. Textbook of Rheumatology 6th ed. Philadelphia: WB Saunders, 2001:55–72.
                    5. Campbell IK, Piccoli DS, Hamilton JA. Stimulation of human chondrocyte prostaglandin E2 production by recombinant human interleukin-1 and tumour necrosis factor.. Biochim Biophys Acta 1990 Mar 9;1051(3):310-8.
                      pubmed: 2310781doi: 10.1016/0167-4889(90)90140-9google scholar: lookup
                    6. Maier R, Bilbe G, Rediske J, Lotz M. Inducible nitric oxide synthase from human articular chondrocytes: cDNA cloning and analysis of mRNA expression.. Biochim Biophys Acta 1994 Sep 21;1208(1):145-50.
                      pubmed: 7522054doi: 10.1016/0167-4838(94)90171-6google scholar: lookup
                    7. Alwan WH, Carter SD, Dixon JB, Bennett D, May SA, Edwards GB. Interleukin-1-like activity in synovial fluids and sera of horses with arthritis.. Res Vet Sci 1991 Jul;51(1):72-7.
                      pubmed: 1896633doi: 10.1016/0034-5288(91)90034-lgoogle scholar: lookup
                    8. Morris EA, McDonald BS, Webb AC, Rosenwasser LJ. Identification of interleukin-1 in equine osteoarthritic joint effusions.. Am J Vet Res 1990 Jan;51(1):59-64.
                      pubmed: 2301820
                    9. Morris EA, Treadwell BV. Effect of interleukin 1 on articular cartilage from young and aged horses and comparison with metabolism of osteoarthritic cartilage.. Am J Vet Res 1994 Jan;55(1):138-46.
                      pubmed: 8141486
                    10. Caron JP, Tardif G, Martel-Pelletier J, DiBattista JA, Geng C, Pelletier JP. Modulation of matrix metalloprotease 13 (collagenase 3) gene expression in equine chondrocytes by interleukin 1 and corticosteroids.. Am J Vet Res 1996 Nov;57(11):1631-4.
                      pubmed: 8915443
                    11. Richardson DW, Dodge GR. Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes.. Am J Vet Res 2000 Jun;61(6):624-30.
                      pubmed: 10850836doi: 10.2460/ajvr.2000.61.624google scholar: lookup
                    12. MacDonald MH, Stover SM, Willits NH, Benton HP. Regulation of matrix metabolism in equine cartilage explant cultures by interleukin 1.. Am J Vet Res 1992 Dec;53(12):2278-85.
                      pubmed: 1476308
                    13. Frisbie DD, Sandler EA, Trotter GW, McIlwraith CW. Metabolic and mitogenic activities of insulin-like growth factor-1 in interleukin-1-conditioned equine cartilage.. Am J Vet Res 2000 Apr;61(4):436-41.
                      pubmed: 10772110doi: 10.2460/ajvr.2000.61.436google scholar: lookup
                    14. Frean SP, Gettinby G, May SA, Lees P. Influence of interleukin-1beta and hyaluronan on proteoglycan release from equine navicular hyaline cartilage and fibrocartilage.. J Vet Pharmacol Ther 2000 Apr;23(2):67-72.
                      pubmed: 10849250doi: 10.1046/j.1365-2885.2000.00247.xgoogle scholar: lookup
                    15. Iqbal J, Dudhia J, Bird JL, Bayliss MT. Age-related effects of TGF-beta on proteoglycan synthesis in equine articular cartilage.. Biochem Biophys Res Commun 2000 Aug 2;274(2):467-71.
                      pubmed: 10913361doi: 10.1006/bbrc.2000.3167google scholar: lookup
                    16. Kato H, Ohashi T, Nakamura N, Nishimura Y, Watari T, Goitsuka R, Tsujimoto H, Hasegawa A. Molecular cloning of equine interleukin-1 alpha and -beta cDNAs.. Vet Immunol Immunopathol 1995 Oct;48(3-4):221-31.
                      pubmed: 8578682doi: 10.1016/0165-2427(95)05441-8google scholar: lookup
                    17. Howard RD, McIlwraith CW, Trotter GW, Nyborg JK. CLoning of equine interleukin 1 alpha and equine interleukin 1 beta and determination of their full-length cDNA sequences.. Am J Vet Res 1998 Jun;59(6):704-11.
                      pubmed: 9622738
                    18. Martel-Pelletier J, McCollum R, DiBattista J, Faure MP, Chin JA, Fournier S, Sarfati M, Pelletier JP. The interleukin-1 receptor in normal and osteoarthritic human articular chondrocytes. Identification as the type I receptor and analysis of binding kinetics and biologic function.. Arthritis Rheum 1992 May;35(5):530-40.
                      pubmed: 1533521doi: 10.1002/art.1780350507google scholar: lookup
                    19. Chin JE, Horuk R. Interleukin 1 receptors on rabbit articular chondrocytes: relationship between biological activity and receptor binding kinetics.. FASEB J 1990 Mar;4(5):1481-7.
                      pubmed: 2137805doi: 10.1096/fasebj.4.5.2137805google scholar: lookup
                    20. DiBattista JA, Martel-Pelletier J, Fujimoto N, Obata K, Zafarullah M, Pelletier JP. Prostaglandins E2 and E1 inhibit cytokine-induced metalloprotease expression in human synovial fibroblasts. Mediation by cyclic-AMP signalling pathway.. Lab Invest 1994 Aug;71(2):270-8.
                      pubmed: 8078306
                    21. May SA, Hooke RE, Lees P. Interleukin-1 stimulation of equine articular cells.. Res Vet Sci 1992 May;52(3):342-8.
                      pubmed: 1620968doi: 10.1016/0034-5288(92)90035-zgoogle scholar: lookup
                    22. May SA, Hooke RE, Lees P. Species restrictions demonstrated by the stimulation of equine cells with recombinant human interleukin-1.. Vet Immunol Immunopathol 1992 Jan 31;30(4):373-84.
                      pubmed: 1546442doi: 10.1016/0165-2427(92)90106-zgoogle scholar: lookup
                    23. Southern EM. Measurement of DNA length by gel electrophoresis.. Anal Biochem 1979 Dec;100(2):319-23.
                      pubmed: 525781doi: 10.1016/0003-2697(79)90235-5google scholar: lookup
                    24. Richardson DW, Dodge GR. Molecular characteristics of equine stromelysin and the tissue inhibitor of metalloproteinase 1.. Am J Vet Res 1998 Dec;59(12):1557-62.
                      pubmed: 9858406
                    25. Fenton JI, Chlebek-Brown KA, Peters TL, Caron JP, Orth MW. Glucosamine HCl reduces equine articular cartilage degradation in explant culture.. Osteoarthritis Cartilage 2000 Jul;8(4):258-65.
                      pubmed: 10903879doi: 10.1053/joca.1999.0299google scholar: lookup
                    26. Chavira R Jr, Burnett TJ, Hageman JH. Assaying proteinases with azocoll.. Anal Biochem 1984 Feb;136(2):446-50.
                      pubmed: 6426343doi: 10.1016/0003-2697(84)90242-2google scholar: lookup
                    27. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids.. Anal Biochem 1982 Oct;126(1):131-8.
                      pubmed: 7181105doi: 10.1016/0003-2697(82)90118-xgoogle scholar: lookup
                    28. Pawate S, Schey KL, Meier GP, Ullian ME, Mais DE, Halushka PV. Expression, characterization, and purification of C-terminally hexahistidine-tagged thromboxane A2 receptors.. J Biol Chem 1998 Aug 28;273(35):22753-60.
                      pubmed: 9712907doi: 10.1074/jbc.273.35.22753google scholar: lookup
                    29. Saklatvala J, Bird T. A common class of receptors for the two types of porcine interleukin-1 on articular chondrocytes.. Lymphokine Res 1986;5 Suppl 1:S99-104.
                      pubmed: 2946906
                    30. Smith RJ, Rohloff NA, Sam LM, Justen JM, Deibel MR, Cornette JC. Recombinant human interleukin-1 alpha and recombinant human interleukin-1 beta stimulate cartilage matrix degradation and inhibit glycosaminoglycan synthesis.. Inflammation 1989 Aug;13(4):367-82.
                      pubmed: 2668162doi: 10.1007/bf00914921google scholar: lookup
                    31. Yamada H, Kikuchi T, Nemoto O, Obata K, Sato H, Seiki M, Shinmei M. Effects of indomethacin on the production of matrix metalloproteinase-3 and tissue inhibitor of metalloproteinases-1 by human articular chondrocytes.. J Rheumatol 1996 Oct;23(10):1739-43.
                      pubmed: 8895151
                    32. Martel-Pelletier J, Zafarullah M, Kodama S, Pelletier JP. In vitro effects of interleukin 1 on the synthesis of metalloproteases, TIMP, plasminogen activators and inhibitors in human articular cartilage.. J Rheumatol Suppl 1991 Feb;27:80-4.
                      pubmed: 1851231
                    33. Shingu M, Nagai Y, Isayama T, Naono T, Nobunaga M, Nagai Y. The effects of cytokines on metalloproteinase inhibitors (TIMP) and collagenase production by human chondrocytes and TIMP production by synovial cells and endothelial cells.. Clin Exp Immunol 1993 Oct;94(1):145-9.
                      pmc: PMC1534355pubmed: 8403497doi: 10.1111/j.1365-2249.1993.tb05992.xgoogle scholar: lookup
                    34. Tsai MJ, O'Malley BW. Molecular mechanisms of action of steroid/thyroid receptor superfamily members.. Annu Rev Biochem 1994;63:451-86.
                      pubmed: 7979245doi: 10.1146/annurev.bi.63.070194.002315google scholar: lookup
                    35. Krane SM. Some molecular mechanisms of glucocorticoid action.. Br J Rheumatol 1993 May;32 Suppl 2:3-5.
                      pubmed: 8495278doi: 10.1093/rheumatology/32.suppl_2.3google scholar: lookup
                    36. Richardson DW, Dodge GR. Dose-dependent effects of corticosteroids on the expression of matrix-related genes in normal and cytokine-treated articular chondrocytes.. Inflamm Res 2003 Jan;52(1):39-49.
                      pubmed: 12608648doi: 10.1007/s000110300012google scholar: lookup
                    37. Amin AR, Attur M, Patel RN, Thakker GD, Marshall PJ, Rediske J, Stuchin SA, Patel IR, Abramson SB. Superinduction of cyclooxygenase-2 activity in human osteoarthritis-affected cartilage. Influence of nitric oxide.. J Clin Invest 1997 Mar 15;99(6):1231-7.
                      pmc: PMC507937pubmed: 9077531doi: 10.1172/jci119280google scholar: lookup
                    38. Ben-Av P, Crofford LJ, Wilder RL, Hla T. Induction of vascular endothelial growth factor expression in synovial fibroblasts by prostaglandin E and interleukin-1: a potential mechanism for inflammatory angiogenesis.. FEBS Lett 1995 Sep 18;372(1):83-7.
                      pubmed: 7556649doi: 10.1016/0014-5793(95)00956-agoogle scholar: lookup
                    39. Pelletier JP, Martel-Pelletier J. Evidence for the involvement of interleukin 1 in human osteoarthritic cartilage degradation: protective effect of NSAID.. J Rheumatol Suppl 1989 Aug;18:19-27.
                      pubmed: 2553967
                    40. Suda T, Udagawa N, Nakamura I, Miyaura C, Takahashi N. Modulation of osteoclast differentiation by local factors.. Bone 1995 Aug;17(2 Suppl):87S-91S.
                      pubmed: 8579904doi: 10.1016/8756-3282(95)00185-ggoogle scholar: lookup
                    41. Järvinen TA, Moilanen T, Järvinen TL, Moilanen E. Nitric oxide mediates interleukin-1 induced inhibition of glycosaminoglycan synthesis in rat articular cartilage.. Mediators Inflamm 1995;4(2):107-11.
                      pmc: PMC2365616pubmed: 18475625doi: 10.1155/s0962935195000184google scholar: lookup
                    42. Amin AR, Di Cesare PE, Vyas P, Attur M, Tzeng E, Billiar TR, Stuchin SA, Abramson SB. The expression and regulation of nitric oxide synthase in human osteoarthritis-affected chondrocytes: evidence for up-regulated neuronal nitric oxide synthase.. J Exp Med 1995 Dec 1;182(6):2097-102.
                      pmc: PMC2192230pubmed: 7500055doi: 10.1084/jem.182.6.2097google scholar: lookup
                    43. Frean SP, Bryant CE, Fröling IL, Elliott J, Lees P. Nitric oxide production by equine articular cells in vitro.. Equine Vet J 1997 Mar;29(2):98-102.
                      pubmed: 9104557doi: 10.1111/j.2042-3306.1997.tb01649.xgoogle scholar: lookup
                    44. Murrell GA, Jang D, Williams RJ. Nitric oxide activates metalloprotease enzymes in articular cartilage.. Biochem Biophys Res Commun 1995 Jan 5;206(1):15-21.
                      pubmed: 7529496doi: 10.1006/bbrc.1995.1003google scholar: lookup
                    45. Pelletier JP, Mineau F, Ranger P, Tardif G, Martel-Pelletier J. The increased synthesis of inducible nitric oxide inhibits IL-1ra synthesis by human articular chondrocytes: possible role in osteoarthritic cartilage degradation.. Osteoarthritis Cartilage 1996 Mar;4(1):77-84.
                      pubmed: 8731398doi: 10.1016/s1063-4584(96)80009-4google scholar: lookup
                    46. Oh M, Fukuda K, Asada S, Yasuda Y, Tanaka S. Concurrent generation of nitric oxide and superoxide inhibits proteoglycan synthesis in bovine articular chondrocytes: involvement of peroxynitrite.. J Rheumatol 1998 Nov;25(11):2169-74.
                      pubmed: 9818660
                    47. Cao M, Westerhausen-Larson A, Niyibizi C, Kavalkovich K, Georgescu HI, Rizzo CF, Hebda PA, Stefanovic-Racic M, Evans CH. Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target.. Biochem J 1997 May 15;324 ( Pt 1)(Pt 1):305-10.
                      pmc: PMC1218431pubmed: 9164871doi: 10.1042/bj3240305google scholar: lookup

                    Citations

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                    1. 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.
                      doi: 10.1186/s12917-019-2170-8pubmed: 31752879google scholar: lookup
                    2. 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.
                      doi: 10.1186/s12917-018-1426-zpubmed: 29554899google scholar: lookup
                    3. Mobasheri A, Henrotin Y, Biesalski HK, Shakibaei M. Scientific evidence and rationale for the development of curcumin and resveratrol as nutraceutricals for joint health. Int J Mol Sci 2012;13(4):4202-4232.
                      doi: 10.3390/ijms13044202pubmed: 22605974google scholar: lookup
                    4. Ando W, Heard BJ, Chung M, Nakamura N, Frank CB, Hart DA. Ovine synovial membrane-derived mesenchymal progenitor cells retain the phenotype of the original tissue that was exposed to in-vivo inflammation: evidence for a suppressed chondrogenic differentiation potential of the cells. Inflamm Res 2012 Jun;61(6):599-608.
                      doi: 10.1007/s00011-012-0450-xpubmed: 22391623google scholar: lookup
                    5. Clutterbuck AL, Smith JR, Allaway D, Harris P, Liddell S, Mobasheri A. High throughput proteomic analysis of the secretome in an explant model of articular cartilage inflammation. J Proteomics 2011 May 1;74(5):704-15.
                      doi: 10.1016/j.jprot.2011.02.017pubmed: 21354348google scholar: lookup
                    6. Martoriati A, Gérard N. Interleukin-1 (IL-1) system gene expression in granulosa cells: kinetics during terminal preovulatory follicle maturation in the mare. Reprod Biol Endocrinol 2003 May 16;1:42.
                      doi: 10.1186/1477-7827-1-42pubmed: 12803652google scholar: lookup
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