high-density lipoproteins downregulate ccl2 production in human fibroblast-like synoviocytes stimulated by urate crystals高密度脂蛋白下调ccl2生产在人类呈synoviocytes刺激尿酸盐晶体.pdfVIP

Scanu et al. Arthritis Research Therapy 2010, 12:R23 /content/12/1/R23 R E S E A R C H A R T I C L E Research article High-density lipoproteins downregulate CCL2 production in human fibroblast-like synoviocytes stimulated by urate crystals 1 1 2 1 3 1 1 Anna Scanu* , Francesca Oliviero , Lyssia Gruaz , Paolo Sfriso , Assunta Pozzuoli , Federica Frezzato , Carlo Agostini , Danielle Burger†2 and Leonardo Punzi†1 Abstract Introduction: To investigate whether monosodium urate (MSU) crystals induce the production of CCL2 (monocyte chemoattractant protein-1; MCP-1) in human fibroblast-like synoviocytes (FLS) and whether this mechanism would be affected by high-density lipoproteins (HDL). Methods: Human FLS isolated from synovial tissue explants were stimulated with MSU crystals (0.01 to 0.5 mg/ml) or interleukin (IL)-1β (10 pg/ml) in the presence or absence of HDL (50 and 100 μg/ml). The production and expression of CCL2 was evaluated with ELISA, confocal microscopy, immunofluorescence microscopy, chemotaxis assay, and real- time quantitative PCR. Results: Exposure of FLS to MSU crystals induced CCL2 accumulation in culture medium in a dose- and time- dependent manner, reaching a plateau at 50 to 75 μg/ml MSU crystals and 20 to 24 hours. Although low, the induced CCL2 levels were sufficient to trigger mononuclear cell migration. In resting FLS, CCL2 was localized in small cytoplasmic vesicles whose number diminished with MSU crystal stimulation. Concomitantly, MSU crystals triggered the induction of CCL2 mRNA expression. All these processes were inhibited by HDL, which cause a 50% decrease in CCL2 mRNA levels and a dose-dependent inhibition of the release of CCL2. Similar results were obtained when FLS were pretreated with HDL and washed before activation by MSU crystals or IL