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  • Title: Comparative effects of IL-1beta and hydrogen peroxide (H2O2) on catabolic and anabolic gene expression in juvenile bovine chondrocytes.
    Author: Martin G, Andriamanalijaona R, Mathy-Hartert M, Henrotin Y, Pujol JP.
    Journal: Osteoarthritis Cartilage; 2005 Oct; 13(10):915-24. PubMed ID: 15950497.
    Abstract:
    OBJECTIVE: To compare the effects of hydrogen peroxide (H(2)O(2)) to those of interleukin-1beta (IL-1beta) on gene expression in juvenile bovine articular chondrocytes (BAC). The study analyses the activation of nuclear factor-kappa B (NF-kappaB) and activator protein-1 (AP-1) transcription factors, and the mRNA steady-state levels of the type II collagen, aggrecan core protein matrix, metalloproteinases (MMP-1, -3), and transforming growth factor-beta1 (TGF-beta1) genes. METHODS: Confluent BAC cultures were treated for 3 and 24h with IL-1beta and/or different concentrations of H(2)O(2) (Protocol 1). Following initial treatment, a part of the cells was further subjected to another 24h with medium, in the presence of IL-1beta, to determine the effect of the cytokine on H(2)O(2) pre-treated cells (Protocol 2). Total RNA and nuclear protein extractions were performed to study mRNA steady-state levels (real-time polymerase chain reaction) and AP-1/NF-kappaB DNA binding (Electrophoretic Mobility Shift Assays), respectively. RESULTS: IL-1beta enhanced both AP-1 and NF-kappaB binding, whereas H(2)O(2) only activated AP-1. H(2)O(2) pre-treatment decreased the IL-1beta activation of NF-kappaB. Both H(2)O(2) and IL-1beta down-regulated type II collagen and aggrecan expression and increased that of MMP-1 and -3. When cells were pre-treated with H(2)O(2), followed by IL-1beta, the effects were the same as those observed with H(2)O(2) alone. However, although H(2)O(2) and IL-1beta were capable of increasing TGF-beta1 expression separately, subsequent incubation with both factors led to a partial or total abolition of TGF-beta1 up-regulation. CONCLUSION: The different regulation of NF-kappaB and AP-1 by H(2)O(2) and IL-1beta underlines the distinct roles played by the two transcription factors in the regulation of gene expression. H(2)O(2) and IL-1beta exert similar effects on matrix, MMPs and TGF-beta1 gene expression. However, the association of H(2)O(2) and IL-1beta does not cause synergic effect, and rather leads, in some cases, to an opposite effect. These data provide further insights into the respective roles of reactive oxygen species and cytokine in the pathophysiology of joint diseases.
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