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Browsing Medical Sciences by Author "A. Elkhider"

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    IL-34 modulates rheumatoid synovial fibroblast proliferation and migration via ERK/AKT signalling pathway
    (CLINICAL AND EXPERIMENTAL RHEUMATOLOGY, 2020) A. Elkhider; J. Wei; M. AL-Azab; Y. Tang; W. Walana; W. Li; B. Yuan; Y. Ye; G. Wang; Y. Zhang; X. Li
    Objective The interface between pro-inflammatory cytokines and rheumatoid synovial fibroblast (sF LS) has central effects on rheumatoid arthritis (RA). The present study aimed to explore the role of IL-34 expression as one of major cytokine implicated in RA. Methods We examined the expression of IL-34 after RA sF LS stimulated by IL-1B and TGF-B1 separately by reverse transcription polymerase chain reaction (RT-PCR). Transwell and wound closure techniques were used to detect whether IL-34 is involved in promoting cell migration. Cellular viability was determined via CCK-8 and cultural morphology assays between IL-34 downregulated group and non-transfected counterpart. We also tested the expression of VEGF gene with RT-PCR analysis and activation of the major signalling pathways by western blot in IL-34 down-regulated group, IL-16 or TGF-{1 treated groups. Propidium iodide (PI) staining and fluoresceine isothiocyanate (FITC) Annexin V and propidium iodide apoptosis assay were used to analyse cell cycle arrest and apoptosis separately in IL-34 down-regulated cells. Results We found that IL-1f significantly enhanced IL-34 expression, while contrarily, TGF -(1 restrained IL-34 gene expression. Transwell and wound closure techniques showed that IL-34 was involved considerably in promoting cell migration. However, IL-34 knock-down restricted sFLS migration possibly through the diminishing of MMP2 and MMP9 expression. Interestingly, IL-34 down-regulated cells exhibited significantly low cellular viability compared with the non-transfected counterpart via CCK-8 and cultural morphology assays. We found that IL-34 down-regulated cells have low VEGF gene expression compared with treated cells. PI staining showed a GO/GI cell cycle arrest in IL-34 down-regulated cells. FITC Annexin V and propidium iodide apoptosis assay verified that IL-34 down-regulated cells induced massive apoptosis through apoptotic signalling caspase3, while IL-1f treated cells presented termination of cellular apoptosis signalled by BCL-2. Furthermore, we observed IL-34 induced activation of ERK1/2 and AKT pathways while IL-34 down-regulation significantly decreased the activation of these pathways.