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Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 1998 Oct;57(10):602–605. doi: 10.1136/ard.57.10.602

Hyaluronan production in human rheumatoid fibroblastic synovial lining cells is increased by interleukin 1β but inhibited by transforming growth factor β1

M Kawakami 1, K Suzuki 1, Y Matsuki 1, T Ishizuka 1, T Hidaka 1, T Konishi 1, M Matsumoto 1, K Kataharada 1, H Nakamura 1
PMCID: PMC1752477  PMID: 9893571

Abstract

OBJECTIVES—To investigate the regulatory roles of interleukin 1β (IL1β), tumour necrosis factor α (TNFα), interferon γ (IFNγ) or transforming growth factor β1 (TGFβ1) on hyaluronan (HA) synthesis by human fibroblastic synovial lining cells.
METHODS—Concentrations of HA in culture supernatants of fibroblastic synovial lining cell line (RAMAK-1 cell line) with or without stimulation by IL1β, TNFα, IFNγ or TGFβ1 were measured by sandwich binding protein assay. Levels of HA synthase mRNA of the cells with or without stimulation were detected by reverse transcribed polymerase chain reaction. Molecular weights of HA in the culture supernatants of the cells with or without stimulation were measured using high performance gel permeation liquid chromatography.
RESULTS—HA synthesis by the cells was not significantly augmented by TNFα or by IFNγ. It was significantly stimulated by IL1β but inhibited by TGFβ1. Molecular weights of HA in the culture supernatants of the cells were unchanged by stimulation with TNFα. They were remarkably increased by stimulation with IL1β and IFNγ, but reduced with TGFβ1.
CONCLUSION—IL1β is an up regulator of HA synthesis, while TGFβ1 is a down regulator. HA production in the synovial lining cells of inflamed joints (for example, rheumatoid arthritis) might be regulated by the balance of these cytokines.

 Keywords: synovial lining cells; hyaluronan, interleukin 1β; transforming growth factor β1

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Figure 1  .

Figure 1  

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HA production by RAMAK-1 cells. RAMAK-1 cells were incubated for 24 or 48 hours in culture dishes with RPMI-1640 medium containing 10% FBS with or without stimulation by IL1β (1 ng/ml), TNFα (300 units/ml), IFNγ (1000 units/ml) or TGFβ1 (1 ng/ml). At the end of culture, the concentration of HA in the supernatants was measured by sandwich binding protein assay.

Figure 2  .

Figure 2  

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Time and dose dependent inhibition of HA production by RAMAK-1 cells cultured with RPMI-1640 medium containing 10% FBS with or without stimulation by TGFβ1. Cells were incubated for up to 96 hours in the presence of TGFβ1. Supernatants were removed for HA determinations by sandwich binding protein assay at 24, 48, 78 or 96 hours. TGFβ1 concentrations indicated were used.

Figure 3  .

Figure 3  

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HA synthase mRNA of RAMAK-1 cells when stimulated for six hours by IL1β, TNFα, IFNγ or TGFβ1 using RT-PCR. Lane 1, without stimulation; lane 2, with stimulation by IL1β; lane 3, with stimulation by TNFα; lane 4, with stimulation by IFNγ; lane 5, with stimulation by TGFβ1. The primer set amplifies a 548-bp sequence from cDNA transcribed from HA synthase mRNA.

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