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TNF-α, IFN-γ, and IL−1β modulate hyaluronan synthase expression in human skin fibroblasts: Synergistic effect by concomital treatment with FeSO4 plus ascorbate

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Abstract

Several reports have shown that a number of cytokines such as tumor necrosis-α (TNF-α), interferon-γ (IFN-γ), and interleukin-β (IL-1β) are capable to induce hyaluronan sinthases (HASs) mRNA expression in different cell culture types. The obvious consequence of this stimulation is a marked increment in hyaluronan (HA) production. It has been also reported that oxidative stress, by itself, may increase HA levels. The aim of this study was to evaluate how TNF-α, IFN-γ,IL−1β, and exposition to oxidative stress may modulate HAS activities in normal human skin fibroblasts. Moreover, the effects on HAS mRNA expression of the concomitant treatment with cytokines and oxidants, and the HA concentrations after treatments, were studied. TNF-α, IFN-γ, and IL-1β were added to normal or/and exposed to FeSO4 plus ascorbate fibroblast cultures and HAS1, HAS2 and HAS3 mRNA content, by PCR-real time, was assayed 3,h later. HA levels were also evaluated after 24,h incubation. The treatment of fibroblasts with cytokines up-regulated HASs gene expression and increased HA production. IL-1β induced HAS mRNA expression and HA production more efficiently than TNF-α and IFN-γ. The exposition of the fibroblasts with the oxidant system markedly increased HAS activities while slightly HA production. The concomitant treatment of cells with the cytokines and the oxidant was able to further enhance, in a dose dependent way, with synergistic effect on HAS mRNA expression. On the contrary HA levels resulted unaffected by the concomitant treatment, and resemble those obtained with the exposure to FeSO4 plus ascorbate only. This lack in HA production could be due to the deleterious action of free radicals on the HA synthesis.

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Abbreviations

GAGs:

glycosaminoglycans

PGs:

proteoglycans

ECM:

extracellular matrix

HA:

hyaluronic acid

HASs:

hyaluronan synthases

TNF-α:

tumor necrosis factor alpha

IFN-γ:

interferon gamma

IL-1β:

interleukin beta

ROS:

reactive oxygen species

DMEM:

Dulbecco’s minimal essential medium

FBS:

foetal bovine serum

NADH:

reduced nicotinamide adenine dinucleotide

EDTA:

ethylenediaminetetraacetic acid

SDS:

sodium dodecylsulphate

PBS:

buffer phosphate saline

S.D.:

standard deviation

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Authors and Affiliations

  1. Department of Biochemical, Physiological and Nutritional Sciences, School of Medicine, University of Messina, Policlinico Universitario, 98125, Messina, Italy

    Giuseppe M. Campo, Angela Avenoso, Salvatore Campo, D'Ascola Angela & Alberto Calatroni

  2. Department of Morphology, Biochemistry, Physiology and Animal Production, School of Veterinary Medicine, University of Messina, contrada Annunziata, 98168, Messina, Italy

    Alida M. Ferlazzo

  3. PhD Department of Biochemical, Physiological and Nutritional Sciences, School of Medicine, University of Messina, Policlinico Universitario, Torre Biologica, 5∘ piano, Via C. Valeria, 98125, Messina, Italy

    Giuseppe M. Campo

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  1. Giuseppe M. Campo
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  2. Angela Avenoso
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  3. Salvatore Campo
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  4. D'Ascola Angela
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  6. Alberto Calatroni
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Correspondence to Giuseppe M. Campo.

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Campo, G.M., Avenoso, A., Campo, S. et al. TNF-α, IFN-γ, and IL−1β modulate hyaluronan synthase expression in human skin fibroblasts: Synergistic effect by concomital treatment with FeSO4 plus ascorbate. Mol Cell Biochem 292, 169–178 (2006). https://doi.org/10.1007/s11010-006-9230-7

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