Resistin induces expression of proinflammatory cytokines and chemokines in human articular chondrocytes via transcription and messenger RNA stabilization

Abstract

Objective To elucidate the effect of resistin on human articular chondrocytes, and generate a picture of their regulation at the transcriptional and post-transcriptional levels. Methods Human articular chondrocytes were cultured with resistin. Changes in gene expression were analyzed at various doses and times. Cells were also treated with the transcriptional inhibitor actinomycin D after resistin treatment, or the nuclear factor kappa B (NF-κB) inhibitor IKK-NBD before resistin treatment. Gene expression was tested by quantitative real-time polymerase chain reaction. Computational analysis for transcription factor binding motifs was performed on the promoter regions of differentially expressed genes. TC28 chondrocytes were transfected with CCL3 and CCL4 promoter constructs, pNF-κB reporter, and NF-κB and CCAAT/enhancer-binding protein β (C/EBPβ) expression vectors with or without resistin. Results Resistin-treated human articular chondrocytes increased the expression of cytokines and chemokines. The mRNAs for MMP-1, MMP-13 and ADAMTS-4 also increased while COL2A1 and aggrecan were down-regulated. Cytokine and chemokine genes could be categorized into three groups according to the pattern of mRNA expression in a 24 h time course. One pattern suggested rapid regulation by mRNA stability. The second and third patterns were consistent with transcriptional regulation. Computational analysis suggested the transcription factors NF-κB and C/EBPβ were involved in the resistin-induced up-regulation. This prediction was confirmed by the co-transfection of NF-κB and C/EBPβ, and IKK-NBD inhibition. Conclusion Resistin has diverse effects on gene expression in human chondrocytes affecting chemokines, cytokines and matrix genes. mRNA stabilization and transcriptional up-regulation are involved in resistin-induced gene expression in human chondrocytes.

Cytokine signature associated with disease severity in chronic fatigue syndrome patients

“…Resistin in humans appears to have a significant proinflammatory role by targeting PBMCs, endothelial cells, smooth muscle cells, platelets (30), and chondrocytes (31) and by increasing the release of IL-1β, IL-6, and TNF-α via the NF-κB pathway (32). Resistin has been reported to be a marker of inflammation in systemic lupus erythematosus (SLE) and Crohn's disease in humans (33).…”

Transcriptional Induction of ADAMTS5 Protein by Nuclear Factor-κB (NF-κB) Family Member RelA/p65 in Chondrocytes during Osteoarthritis Development

“…Because this protection was not seen in the ADAMTS4 knockout mice (5), ADAMTS5 is thought to be substantially more important than ADAMTS4 for OA development (6). Although ADAMTS4 expression has been reported to be induced by transforming growth factor-␤ (TGF-␤) and proinflammatory cytokines in chondrocytes, less is known about the regulation of ADAMTS5 expression (7)(8)(9)(10)(11). Despite the substantial information about the expression profiles and the substrates of ADAMTS5 (1), little is known about the upstream signaling or the functional regulation of the ADAMTS5 promoter.…”

CCAAT/Enhancer-binding Protein β and NF-κB Mediate High Level Expression of Chemokine Genes CCL3 and CCL4 by Human Chondrocytes in Response to IL-1β*

“…It can be expected that this increase in a wide range of chemokines will have a significant impact on the cells of cartilage and other related joint tissues and should be considered in the pathophysiology of OA. Recently, we demonstrated that the adipokine, resistin, present in injured joints (12), also increased chemokine genes at both the transcriptional and post-transcriptional levels (13) in temporal patterns similar to the IL-1␤ patterns (6). The post-transcriptional regulation of chemokines was further investigated by us in chondrocytes with resistin (13) and by others in fibroblasts (14).…”