Production of tissue inhibitor of metalloproteinases 3 is selectively enhanced by calcium pentosan polysulfate in human rheumatoid synovial fibroblasts

Masayuki Takizawa, Eiko Ohuchi, Hajime Yamanaka, Hiroyuki Nakamura, Eiji Ikeda, Peter Ghosh, Yasunori Okada

Research output: Contribution to journalArticle

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Abstract

Objective. To determine the effects of calcium pentosan polysulfate (CaPPS) on the production of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMP), in cultures of rheumatoid synovial fibroblasts. Methods. The production of MMP-1, -2, -3, -7, -8, -9, and -13 and of TIMP-1, -2, -3, and -4 in cultured rheumatoid synovial fibroblasts treated with 0.1, 1, and 10 μg/ml CaPPS in the presence and absence of 100 units/ml interleukin-1α (IL-1α) was examined by a sandwich enzyme immunoassay system and/or immuno-blotting. The messenger RNA (mRNA) expression of TIMP-3 and membrane type 1 MMP was determined by Northern blotting, and the cells expressing TIMP-3 gene in rheumatoid synovium were identified by in situ hybridization. The synthesis and secretion of TIMP-3 protein were monitored by pulse-chase experiments. TIMP-3 was immunolocalized in untreated or CaPPS-treated rheumatoid synovial fibroblasts and synovium using an avidin-biotin-peroxidase complex method. Results. Treatment of cultured rheumatoid synovial fibroblasts with CaPPS resulted in a dose-dependent increase in the production of TIMP-3 in both cell lysates and media from the treated cells. However, CaPPS did not affect the levels of the other MMPs or TIMPs examined. The production of TIMP-3 was further enhanced in the cells treated with both IL-1α and CaPPS. Immunohistochemistry confirmed the enhanced production of TIMP-3 by cells exposed to CaPPS. The mRNA level of TIMP-3 increased 3.4-fold by treating rheumatoid synovial fibroblasts with IL-1α, but CaPPS itself did not alter the expression levels in the IL-1α-treated or -untreated cells. Pulse-chase studies demonstrated that translation for TIMP-3 protein was enhanced by CaPPS treatment. In situ hybridization and immunohistochemistry indicated that TIMP-3 was expressed mainly in the hyperplastic lining cells of rheumatoid synovium, and that the production of this protein by these immunoreactive lining cells was significantly increased by treatment with CaPPS. Conclusion. The present study is the first to demonstrate that the new antiarthritic drug, CaPPS, selectively enhanced TIMP-3 production at the posttranscription level in cultured rheumatoid synovial fibroblasts and in the lining cells of rheumatoid synovium. By this mechanism, CaPPS may be able to modulate joint tissue destruction in rheumatoid arthritis.

Original languageEnglish
Pages (from-to)812-820
Number of pages9
JournalArthritis and Rheumatism
Volume43
Issue number4
DOIs
Publication statusPublished - 2000

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Pentosan Sulfuric Polyester
Tissue Inhibitor of Metalloproteinase-3
Fibroblasts
Calcium
Synovial Membrane
Interleukin-1
Matrix Metalloproteinases
In Situ Hybridization
Immunohistochemistry
Matrix Metalloproteinase 14
Tissue Inhibitor of Metalloproteinases
Tissue Inhibitor of Metalloproteinase-2
Messenger RNA
Matrix Metalloproteinase 1
Proteins
Tissue Inhibitor of Metalloproteinase-1
Avidin
Matrix Metalloproteinase 2
Biotin
Immunoenzyme Techniques

ASJC Scopus subject areas

  • Immunology
  • Rheumatology
  • Immunology and Allergy
  • Pharmacology (medical)

Cite this

Production of tissue inhibitor of metalloproteinases 3 is selectively enhanced by calcium pentosan polysulfate in human rheumatoid synovial fibroblasts. / Takizawa, Masayuki; Ohuchi, Eiko; Yamanaka, Hajime; Nakamura, Hiroyuki; Ikeda, Eiji; Ghosh, Peter; Okada, Yasunori.

In: Arthritis and Rheumatism, Vol. 43, No. 4, 2000, p. 812-820.

Research output: Contribution to journalArticle

Takizawa, Masayuki ; Ohuchi, Eiko ; Yamanaka, Hajime ; Nakamura, Hiroyuki ; Ikeda, Eiji ; Ghosh, Peter ; Okada, Yasunori. / Production of tissue inhibitor of metalloproteinases 3 is selectively enhanced by calcium pentosan polysulfate in human rheumatoid synovial fibroblasts. In: Arthritis and Rheumatism. 2000 ; Vol. 43, No. 4. pp. 812-820.
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T1 - Production of tissue inhibitor of metalloproteinases 3 is selectively enhanced by calcium pentosan polysulfate in human rheumatoid synovial fibroblasts

AU - Takizawa, Masayuki

AU - Ohuchi, Eiko

AU - Yamanaka, Hajime

AU - Nakamura, Hiroyuki

AU - Ikeda, Eiji

AU - Ghosh, Peter

AU - Okada, Yasunori

PY - 2000

Y1 - 2000

N2 - Objective. To determine the effects of calcium pentosan polysulfate (CaPPS) on the production of matrix metalloproteinases (MMPs) and their endogenous inhibitors, tissue inhibitors of metalloproteinases (TIMP), in cultures of rheumatoid synovial fibroblasts. Methods. The production of MMP-1, -2, -3, -7, -8, -9, and -13 and of TIMP-1, -2, -3, and -4 in cultured rheumatoid synovial fibroblasts treated with 0.1, 1, and 10 μg/ml CaPPS in the presence and absence of 100 units/ml interleukin-1α (IL-1α) was examined by a sandwich enzyme immunoassay system and/or immuno-blotting. The messenger RNA (mRNA) expression of TIMP-3 and membrane type 1 MMP was determined by Northern blotting, and the cells expressing TIMP-3 gene in rheumatoid synovium were identified by in situ hybridization. The synthesis and secretion of TIMP-3 protein were monitored by pulse-chase experiments. TIMP-3 was immunolocalized in untreated or CaPPS-treated rheumatoid synovial fibroblasts and synovium using an avidin-biotin-peroxidase complex method. Results. Treatment of cultured rheumatoid synovial fibroblasts with CaPPS resulted in a dose-dependent increase in the production of TIMP-3 in both cell lysates and media from the treated cells. However, CaPPS did not affect the levels of the other MMPs or TIMPs examined. The production of TIMP-3 was further enhanced in the cells treated with both IL-1α and CaPPS. Immunohistochemistry confirmed the enhanced production of TIMP-3 by cells exposed to CaPPS. The mRNA level of TIMP-3 increased 3.4-fold by treating rheumatoid synovial fibroblasts with IL-1α, but CaPPS itself did not alter the expression levels in the IL-1α-treated or -untreated cells. Pulse-chase studies demonstrated that translation for TIMP-3 protein was enhanced by CaPPS treatment. In situ hybridization and immunohistochemistry indicated that TIMP-3 was expressed mainly in the hyperplastic lining cells of rheumatoid synovium, and that the production of this protein by these immunoreactive lining cells was significantly increased by treatment with CaPPS. Conclusion. The present study is the first to demonstrate that the new antiarthritic drug, CaPPS, selectively enhanced TIMP-3 production at the posttranscription level in cultured rheumatoid synovial fibroblasts and in the lining cells of rheumatoid synovium. By this mechanism, CaPPS may be able to modulate joint tissue destruction in rheumatoid arthritis.

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