TNF-α/calreticulin dual signaling induced NLRP3 inflammasome activation associated with HuR nucleocytoplasmic shuttling in rheumatoid arthritis
| Autor: | Wei Wei, Xuguo Sun, Jun Ma, Chunyou Wan, Yixin Liu, Yingyu Bai, Yang Wang, Fang Zheng |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
musculoskeletal diseases
0301 basic medicine Inflammasomes Immunology ELAV-Like Protein 1 Arthritis Rheumatoid 03 medical and health sciences 0302 clinical medicine Western blot NLR Family Pyrin Domain-Containing 3 Protein medicine Human Umbilical Vein Endothelial Cells Gene silencing Humans Secretion Pharmacology Gene knockdown biology medicine.diagnostic_test Chemistry Tumor Necrosis Factor-alpha Synovial Membrane Signal transducing adaptor protein Inflammasome Molecular biology Synoviocytes 030104 developmental biology Apoptosis biology.protein Calreticulin 030215 immunology medicine.drug Signal Transduction |
| Zdroj: | Inflammation research : official journal of the European Histamine Research Society ... [et al.]. 68(7) |
| ISSN: | 1420-908X |
| Popis: | The present study was undertaken to validate whether TNF-α and calreticulin (CRT) serve as dual signaling to activate nucleotide-binding oligomerization domain-, leucine-rich repeat- and pyrin domain-containing 3 (NLRP3) inflammasome in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS) and HUVECs. The effect of human antigen R (HuR) in NLRP3 inflammasome activation was also explored in RA FLS. Immunofluorescence was used to determine the expression of NLRP3 and adaptor protein apoptosis associated speck-like protein containing a CARD (ASC) in RA synovial tissue and HuR location in RA FLS. Western blot and quantitative real-time PCR were employed to measure the priming effect of NLRP3 inflammasome in cells and HuR expression in synovial tissue. The concentrations of IL-1β and IL-18 were detected by enzyme linked immunosorbent assay. Immunohistochemistry was used to visualize the expression of HuR in synovial tissue. HuR knockdown in RA FLS was achieved by siRNA-mediated gene silencing. Higher expression of NLRP3 and ASC in RA synovial tissue than those in osteoarthritis was detected. The staining of NLRP3, ASC and cleaved IL-1β were observed in FLS and vascular endothelial cells in RA synovium. Expression of NLRP3 and pro-IL-1β in RA FLS and HUVECs treated with TNF-α was increased. The pro-IL-18 expression was also enhanced in HUVECs, but not in RA FLS. TNF-α/CRT dual stimulation of cells gave rise to caspase-1 p20 expression and the secretion of IL-1β. The secreted IL-18 was also elevated in HUVECs but not in RA FLS. HuR expression was significantly elevated in RA synovial tissue. TNF-α initiated the nucleocytoplasmic shuttling of HuR in both FLS and HUVECs. The knockdown of HuR in FLS incubated with TNF-α led to reduced caspase-1 p20 protein expression and further resulted in decreased secretion of IL-1β in the presence of CRT. TNF-α/CRT dual signaling induced NLRP3 inflammasome activation, which could be suppressed by HuR knockdown presumably due to the block of HuR translocating from nucleus to cytoplasma. |
| Databáze: | OpenAIRE |
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