Hypoxia Differentially Affects Il-1 Beta-Stimulated Mmp-1 And Mmp-13 Expression Of Fibroblast-Like Synoviocytes In An Hif-1 Alpha-Dependent Manner

Objectives. To further understand the expression regulation of MMP-1 and MMP-13 under physiological and pathological conditions, we investigated the combined effects of hypoxia and pro-inflammatory stimuli on the expression of MMP-1 and MMP-13 in rheumatoid synovial fibroblasts.Methods. Synovial fibroblasts were cultured under either hypoxic or normoxic conditions in the presence of IL-1 beta stimulation. The culture supernatant was analysed for secreted levels of VEGF, MMP-1 and MMP-13. Their gene expression was quantified with real-time and semi-quantitative PCR. Another group of cells was transfected with small-interfering RNA (siRNA) specific for hypoxia-inducible factor-1 alpha (HIF-1 alpha). The protein levels of HIF-1 alpha were detected by western blot analysis.Results. In response to 10 ng/ml of IL-1 beta under normoxia, the levels of MMP-1 and MMP-13 increased compared with the levels observed under hypoxia. IL-1 beta stimulation under hypoxia induced a 2-fold increase in the level of MMP-1 and a 2-fold decrease in the level of MMP-13 compared with cells cultured under normoxia. A similar pattern of differential expression for MMP-1 and MMP-13 was observed with 1 and 5 ng/ml IL-1 beta, but not at 0.1 ng/ml. The differential expression of MMPs under the combined effect of IL-1 beta and hypoxia was significantly attenuated by silencing HIF-1 alpha with siRNA.Conclusions. Hypoxia in arthritic joints may differentially affect the IL-1 beta-stimulated expression of MMP-1 and MMP-13 in rheumatoid synovial fibroblasts. This effect is dependent on HIF-1 alpha expression. This hypoxia-mediated differential effect should be taken into consideration when testing the efficiency of therapies that target HIF-1 alpha.