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Báo cáo y học: "Transcriptional regulation of matrix metalloproteinase-1 and collagen 1A2 explains the anti-fibrotic effect exerted by proteasome inhibition in human dermal fibroblasts"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học General Psychiatry cung cấp cho các bạn kiến thức về ngành y đề tài: Transcriptional regulation of matrix metalloproteinase-1 and collagen 1A2 explains the anti-fibrotic effect exerted by proteasome inhibition in human dermal fibroblasts. | Goffin et al. Arthritis Research Therapy 2010 12 R73 http arthritis-research.eom content 12 2 R73 RESEARCH ARTICLE Open Access Transcriptional regulation of matrix metalloproteinase-1 and collagen 1A2 explains the anti-fibrotic effect exerted by proteasome inhibition in human dermal fibroblasts Laurence Goffin 1 Queralt Seguin-Estevez2 Montserrat Alvarez1 Walter Reith2 and Carlo Chizzolini 1 Abstract Introduction Extracellular matrix ECM turnover is controlled by the synthetic rate of matrix proteins including type I collagen and their enzymatic degradation by matrix metalloproteinases MMPs . Fibrosis is characterized by an unbalanced accumulation of ECM leading to organ dysfunction as observed in systemic sclerosis. We previously reported that proteasome inhibition PI in vitro decreases type I collagen and enhances MMP-1 production by human fibroblasts thus favoring an antifibrotic fibroblast phenotype. These effects were dominant over the pro-fibrotic phenotype induced by transforming growth factor TGF -p. Here we investigate the molecular events responsible for the anti-fibrotic phenotype induced in fibroblasts by the proteasome inhibitor bortezomib. Methods The steady-state mRNA levels of COL1A1 COL1A2 TIMP-1 MMP-1 and MMP-2 were assessed by quantitative PCR in human dermal fibroblasts cultured in the presence of TGF-p bortezomib or both. Transient fibroblast transfection was performed with wild-type and mutated COL1A1 and MMP-1 promoters. Chromatin immunoprecipitation electrophoretic mobility shift assay EMSA and DNA pull-down assays were used to assess the binding of c-Jun SP1 AP2 and Smad2 transcription factors. Immunoblotting and immunofluorescent microscopy were performed for identifying phosphorylated transcription factors and their cellular localization. Results Bortezomib decreased the steady-state mRNA levels of COL1A1 and COL1A2 and abrogated SP1 binding to the promoter of COL1A2 in both untreated and TGF-p-activated fibroblasts. Reduced COL1A2 .