Polysulphated glycosaminoglycans modulate transcription of interleukin-1β treated chondrocytes in monolayer culture

W. D. Mertens; J. N. MacLeod; S. L. Fubini; M. Vernier-Singer; A. J. Nixon; R. J. Todhunter

doi:10.1055/s-0038-1632759

Veterinary and Comparative Orthopaedics and Traumatology, Inhaltsverzeichnis

Vet Comp Orthop Traumatol 2003; 16(02): 93-98
DOI: 10.1055/s-0038-1632759

Original Research

Schattauer GmbH

Polysulphated glycosaminoglycans modulate transcription of interleukin-1β treated chondrocytes in monolayer culture

W. D. Mertens^*

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

,

J. N. MacLeod

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

,

S. L. Fubini

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

,

M. Vernier-Singer

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

,

A. J. Nixon

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

,

R. J. Todhunter

¹College of Veterinary Medicine and the James A. Baker Institute for Animal Health, Cornell University, Ithaca, NY 14853

› Institutsangaben

Abstract

Summary

The ability of polysulphated glycosaminoglycans (PSGAGs, Adequan®) to modulate the transcription of major articular cartilage matrix proteins and enzymes was examined. Northern blot analyses were used to compare steady-state mRNA levels of type-II procollagen, aggrecan core protein, matrix metalloproteinase (MMP)-1, MMP-3, tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-3 in equine chondrocytes grown in monolayer culture. The groups included: control, low-dose (0.1mg/ml) Adequan®, high-dose (1.0mg/ml) Adequan®, IL-1 β (10ng/ml), IL-1 β with low-dose Adequan®, and IL-1 β with high-dose Adequan®. The responses (p<0.05) to treatment were compared with one-way analysis of variance and post hoc least significant difference test. Interleukin-1 β significantly decreased steady-state levels of type-II procollagen (0.5x) and aggrecan core protein (to undetectable levels) and significantly increased MMP-1 (11.7x), MMP-3 (1.8x), TIMP-1 (1.9x), and TIMP-3 (5.1 x). Adequan® alone significantly increased steady-state mRNA levels of type-II procollagen (1.6x) and decreased MMP-1 (0.3x at high-dose only), MMP-3 (0.5x), and TIMP-1 (0.3x at high-dose only). In IL-1 β-treated chondrocytes, Adequan^® significantly increased steady-state mRNA levels of type-II procollagen (1.4x of IL-1 β-treated levels), aggrecan core protein (IL-1 β -treated had no detectable expression), and MMP-3 (3.6 x at high dose) and significantly decreased steady-state mRNA levels of MMP-1 (0.3 x), and TIMP-1 (0.2x). This study is the first to demonstrate that Adequan^® alters steady-state mRNA levels of key matrix proteins and enzymes; that it counteracts some of the deleterious effects of IL-1 β; and that these effects are in part dose-dependent.

Keywords

Chondrocyte - mRNA - polysulphated glycosaminoglycan - interleukin-1 - equine

Volltext

Referenzen

References
1 Adam M. Influence of Arteparon on Disturbances of Collagen and Proteoglycan Metabolism in Osteoarthritic Cartilage. Proceedings. 1-1-1979;9th Eur Cong Rheumatol: 30-8
2 Adam M, Krabcova M, Musilova J, Pesakova V, Brettschneider I, Deyl Z. Contribution to the Mode of Action of Glycosaminoglycan-Polysulphate (GAGPS) Upon Human Osteoarthrotic Cartilage. Biochemical Study of the Collagen and Proteoglycan Turnover. Arzneimittelforschung 1980; 30 (10) 1730-2.
3 Busch S. J., Martin G. A., Barnhart R. L., Mano M, Cardin A. D., Jackson R. L. Trans-Repressor Activity of Nuclear Glycosaminoglycans on Fos and Jun/AP-1 Oncoprotein-Mediated Transcription. J. Cell Biol 1992; 116 (01) 31-42.
4 Chandrasekhar S, Harvey A. K., Higginbotham J. D., Horton W. E. Interleukin-1-Induced Suppression of Type II Collagen Gene Transcription Involves DNA Regulatory Elements. Exp. Cell Res 1990; 191 (01) 105-14.
5 Chomczynski P. A Reagent for the Single-Step Simultaneous Isolation of RNA, DNA and Proteins From Cell and Tissue Samples. Biotechniques 1993; 15 (03) 532-7.
6 Chomczynski P, Mackey K. Substitution of Chloroform by Bromo-Chloropropane in the Single-Step Method of RNA Isolation. Anal. Biochem. 2-10-1995; 225. 01 163-4.
7 Cook J. L., Anderson C. C., Kreeger J. M., Tomlinson J. L. Effects of Human Recombinant Interleukin-1beta on Canine Articular Chondrocytes in Three-Dimensional Culture. Am. J. Vet. Res 2000; 61 (07) 766-70.
8 DiPasquale G, Caccese R, Pasternak R, Conaty J, Hubbs S, Perry K. Proteoglycanand Collagen-Degrading Enzymes From Human Interleukin 1-Stimulated Chondrocytes From Several Species: Proteoglycanase and Collagenase Inhibitors As Potentially New Disease-Modifying Antiarthritic Agents. Proc. Soc. Exp. Biol. Med 1986; 183 (02) 262-7.
9 Fedarko N. S., Conrad H. E. A Unique Heparan Sulfate in the Nuclei of Hepatocytes: Structural Changes With the Growth State of the Cells. J. Cell Biol 1986; 102 (02) 587-99.
10 Fedarko N. S., Ishihara M, Conrad H. E. Control of Cell Division in Hepatoma Cells by Exogenous Heparan Sulfate Proteoglycan. J. Cell Physiol 1989; 139 (02) 287-94.
11 Ghosh P. Chondroprotective Drugs and Osteoarthritis. Ann. Rheum. Dis 1990; 49 (05) 338-9.
12 Glade M. J. Polysulfated Glycosaminoglycan Accelerates Net Synthesis of Collagen and Glycosaminoglycans by Arthritic Equine Cartilage Tissues and Chondrocytes. Am. J. Vet. Res 1990; 51 (05) 779-85.
13 Hering T. M., Kollar J, Huynh T. D., Varelas J. B., Sandell L. J. Modulation of Extracellular Matrix Gene Expression in Bovine High-Density Chondrocyte Cultures by Ascorbic Acid and Enzymatic Resuspension. Arch. Biochem. Biophys 1994; 314 (01) 90-8.
14 Hubbard J. R., Steinberg J. J., Bednar M. S., Sledge C. B. Effect of Purified Human Interleukin-1 on Cartilage Degradation. J. Orthop. Res 1988; 06 (02) 180-7.
15 Ishihara M, Fedarko N. S., Conrad H. E. Transport of Heparan Sulfate into the Nuclei of Hepatocytes. J. Biol. Chem 10-15-1986; 261 (29) 13575-80.
16 Lefebvre V, Garofalo S, Zhou G, Metsaranta M, Vuorio E, de Crombrugghe B. Characterization of Primary Cultures of Chondrocytes From Type II Collagen/Beta-Galactosidase Transgenic Mice. Matrix Biol 1994; 14 (04) 329-35.
17 MacLeod J. N., Fubini S. L., Gu D. N., Tetreault J. W., Todhunter R. J. Effect of Synovitis and Corticosteroids on Transcription of Cartilage Matrix Proteins. Am. J. Vet. Res 1998; 59 (08) 1021-6.
18 McGuire-Goldring M. B., Meats J. E., Wood D. D., Ihrie E. J., Ebsworth N. M., Russell R. G. In Vitro Activation of Human Chondrocytes and Synoviocytes by a Human Interleukin-1-Like Factor. Arthritis Rheum 1984; 27 (06) 654-62.
19 McIlwraith C. W., Vachon A. Review of Pathogenesis and Treatment of Degenerative Joint Disease. Equine Vet. J. Suppl 1988; (06) 3-11.
20 Morris E. A., Treadwell B. V. Effect of Interleukin 1 on Articular Cartilage From Young and Aged Horses and Comparison With Metabolism of Osteoarthritic Cartilage. Am. J. Vet. Res 1994; 55 (01) 138-46.
21 Morris E. A., Wilcon S, Treadwell B. V. Inhibition of Interleukin 1-Mediated Proteoglycan Degradation in Bovine Articular Cartilage Explants by Addition of Sodium Hyaluronate. Am. J. Vet. Res 1992; 53 (11) 1977-82.
22 Nixon A. J., Lust G, Vernier-Singer M. Isolation, Propagation, and Cryopreservation of Equine Articular Chondrocytes. Am. J. Vet. Res 1992; 53 (12) 2364-70.
23 Pasternak R. D., Hubbs S. J., Caccese R. G., Marks R. L., Conaty J. M., DiPasquale G. Interleukin-1 Stimulates the Secretion of Proteoglycan-and Collagen-Degrading Proteases by Rabbit Articular Chondrocytes. Clin. Immunol. Immunopathol 1986; 41 (03) 351-67.
24 Pasternak R. D., Hubbs S. J., Caccese R. G., Marks R. L., Conaty J. M., DiPasquale G. Interleukin-1 Induces Chondrocyte Protease Production: the Development of Collagenase Inhibitors. Agents Actions 1987; 21 (3-4): 328-30.
25 Pelletier J. P., Caron J. P., Evans C, Robbins P. D., Georgescu H. I., Jovanovic D, Fernandes J. C., Martel-Pelletier J. In Vivo Suppression of Early Experimental Osteoarthritis by Interleukin-1 Receptor Antagonist Using Gene Therapy. Arthritis Rheum 1997; 40 (06) 1012-9.
26 Richardson D. W., Dodge G. R. 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; 61 (06) 624-30.
27 Sambrook J, Fritsch E. F., Maniatis T. Molecular Cloning: A Laboratory Manual. 2 ed. Cold Spring Harbor. N. Y: Cold Spring Harbor Laboratory; 1-15-1989.
28 Sevalla K, Todhunter R. J., Vernier-Singer M, Budsberg S. C. Effect of Polysulfated Glycosaminoglycan on DNA Content and Proteoglycan Metabolism in Normal and Osteoarthritic Canine Articular Cartilage Explants. Vet. Surg 2000; 29 (05) 407-14.
29 Sobel M, Fish W. R., Toma N, Luo S, Bird K, Mori K, Kusumoto S, Blystone S. D., Suda Y. Heparin Modulates Integrin Function in Human Platelets. J. Vasc. Surg 2001; 33 (03) 587-94.
30 Steinmeyer J, Burton-Wurster N, Lust G. Effects of Three Antiarthritic Drugs on Fibronectin and Keratan Sulfate Synthesis by Cultured Canine Articular Cartilage Chondrocytes. Am. J. Vet. Res 1992; 53 (11) 2077-83.
31 Stewart M. C., Saunders K. M., Burton-Wurster N, MacLeod J. N. Phenotypic Stability of Articular Chondrocytes in Vitro: the Effects of Culture Models, Bone Morphogenetic Protein 2, and Serum Supplementation. J. Bone Miner. Res 2000; 15 (01) 166-74.
32 Suda Y, Mori K, Bird K, Marques D, Ormsby J, Tanaka S, Koshida S, Nakamura M, Kusumoto S, Sobel M. A Novel Crosslinking Reagent and Its Application for the Detection and Isolation of Heparin-Binding Protein(s) on the Platelet Surface. J. Biochem. (Tokyo) 1999; 125 (06) 1011-5.
33 Takizawa M, Ohuchi E, Yamanaka H, Nakamura H, Ikeda E, Ghosh P, Okada Y. Production of Tissue Inhibitor of Metalloproteinases 3 Is Selectively Enhanced by Calcium Pentosan Polysulfate in Human Rheumatoid Synovial Fibroblasts. Arthritis Rheum 2000; 43 (04) 812-20.
34 Todhunter R. J., Lust G. Polysulfated Glycosaminoglycan in the Treatment of Osteoarthritis. J. Am. Vet. Med. Assoc 4-15-1994; 204 (08) 1245-51.
35 Todhunter R. J., Minor R. R., Wootton J. A., Krook L, Burton-Wurster N, Lust G. Effects of Exercise and Polysulfated Glycosaminoglycan on Repair of Articular Cartilage Defects in the Equine Carpus. J. Orthop. Res 1993; 11 (06) 782-95.
36 Trotter G. W. Polysulfated glycosaminoglycan (adequan). C. Wayne McIlwraith and Gayle W. Trotter. Joint disease in the horse. 1st ed. Philadelphia: W. B. Saunders Co; 1-15-1996. 270-80.
37 Vacha J, Pesakova V, Krajickova J, Adam M. Effect of Glycosaminoglycan Polysulphate on the Metabolism of Cartilage Ribonucleic Acid. Arzneimittelforschung 1984; 34 (05) 607-9.
38 Verbruggen G, Cornelissen M, Elewaut D, Broddelez C, de Ridder L, Veys E. M. Influence of Polysulfated Polysaccharides on Aggrecans Synthesized by Differentiated Human Articular Chondrocytes. J. Rheumatol 1999; 26 (08) 1663-71.
39 Yasuda T, Poole A. R. A Fibronectin Fragment Induces Type II Collagen Degradation by Collagenase Through an Interleukin-1-Mediated Pathway. Arthritis Rheum 2002; 46 (01) 138-48.
40 Yodlowski M. L., Hubbard J. R., Kispert J, Keller K, Sledge C. B., Steinberg J. J. Antibody to Interleukin 1 Inhibits the Cartilage Degradative and Thymocyte Proliferative Actions of Rheumatoid Synovial Culture Medium. J. Rheumatol 1990; 17 (12) 1600-7.