Identification of Novel Chondroprotective Mediators in Resolving Inflammatory Exudates
| Autor: | Karin V. Greco, Trinidad Montero-Melendez, Prashant Mori, Kevin Greenslade, Sarah E. Headland, Costantino Pitzalis, Adrian Moore, Magdalena Kaneva, Mauro Perretti |
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| Rok vydání: | 2017 |
| Předmět: |
Male
Proteomics 0301 basic medicine Inflammatory arthritis Immunology Inflammation Real-Time Polymerase Chain Reaction Mass Spectrometry Glycosaminoglycan Mice 03 medical and health sciences Chondrocytes 0302 clinical medicine In vivo medicine Animals Immunology and Allergy Rats Wistar Pleurisy Aggrecan Catabolism business.industry Cartilage Hemopexin Exudates and Transudates Osteoarthritis Knee medicine.disease Arthritis Experimental Molecular biology Rats Mice Inbred C57BL Disease Models Animal 030104 developmental biology medicine.anatomical_structure 030220 oncology & carcinogenesis medicine.symptom business |
| Zdroj: | The Journal of Immunology. 198:2876-2885 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.1601111 |
| Popis: | We hypothesized that exudates collected at the beginning of the resolution phase of inflammation might be enriched for tissue protective molecules; thus an integrated cellular and molecular approach was applied to identify novel chondroprotective bioactions. Exudates were collected 6 h (inflammatory) and 24 h (resolving) following carrageenan-induced pleurisy in rats. The resolving exudate was subjected to gel filtration chromatography followed by proteomics, identifying 61 proteins. Fractions were added to C28/I2 chondrocytes, grown in micromasses, ions with or without IL-1β or osteoarthritic synovial fluids for 48 h. Three proteins were selected from the proteomic analysis, α1-antitrypsin (AAT), hemopexin (HX), and gelsolin (GSN), and tested against catabolic stimulation for their effects on glycosaminoglycan deposition as assessed by Alcian blue staining, and gene expression of key anabolic proteins by real-time PCR. In an in vivo model of inflammatory arthritis, cartilage integrity was determined histologically 48 h after intra-articular injection of AAT or GSN. The resolving exudate displayed protective activities on chondrocytes, using multiple readouts: these effects were retained in low m.w. fractions of the exudate (46.7% increase in glycosaminoglycan deposition; ∼20% upregulation of COL2A1 and aggrecan mRNA expression), which reversed the effect of IL-1β. Exogenous administration of HX, GSN, or AAT abrogated the effects of IL-1β and osteoarthritic synovial fluids on anabolic gene expression and increased glycosaminoglycan deposition. Intra-articular injection of AAT or GSN protected cartilage integrity in mice with inflammatory arthritis. In summary, the strategy for identification of novel chondroprotective activities in resolving exudates identified HX, GSN and AAT as potential leads for new drug discovery programs. |
| Databáze: | OpenAIRE |
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