Feprazone Ameliorates TNF-α-Induced Loss of Aggrecan via Inhibition of the SOX-4/ADAMTS-5 Signaling Pathway

Abstract

Background: Arthritis is a cartilage degenerative disease that is mainly induced by the degradation of the cartilage extracellular matrix (ECM), which is found to be regulated by the expression level of a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMT-5), an enzyme degrading Aggrecans in the ECM. Feprazone is a classic nonsteroidal anti-inflammatory drug with promising efficacy in arthritis. The present study aims to investigate the protective effect of Feprazone on the degraded Aggrecan in the human chondrocytes induced with tumor necrosis factor-α (TNF-α) and to clarify the underlying mechanism. Methods: To investigate the effect of Feprazone, the CHON-001 chondrocytes were stimulated with TNF-α (10 ng/mL) in the presence or absence of Feprazone (3, 6 μM) for 24 h. Mitochondrial membrane potential was evaluated using the Rhodamine 123 assay. The gene expressions of interleukin-1β (IL-1β), interleukin-8 (IL-8), monocyte chemotactic protein 1 (MCP-1), and ADAMTS-5 in the treated chondrocytes were detected using real-time quantitative polymerase chain reaction (qRT-PCR), and the protein levels of these targets were determined using enzyme-linked immunosorbent assay (ELISA). SOX-4 was knocked down by transfecting the siRNA into the chondrocytes. Western blot analysis was utilized to evaluate the expression levels of SOX-4, Aggrecan, and protein kinase C (PKCα). Results: First, the reduced mitochondrial membrane potential (ΔΨm) and secretion of proinflammatory factors (IL-1β, IL-8, and MCP-1) induced by TNF-α were significantly reversed by treatment with Feprazone. Second, the expression of Aggrecan was significantly decreased by stimulation with TNF-α via upregulation of ADAMTS-5 but was dramatically reversed by the introduction of Feprazone. Third, we found that TNF-α elevated the expression of ADAMTS-5 by upregulating SOX-4, which was observed to be related to the activation of PKCα. Lastly, the elevated expression of SOX-4 induced by TNF-α was significantly reversed by Feprazone. Conclusions: Feprazone might ameliorate TNF-α-induced loss of Aggrecan via the inhibition of the SOX-4/ADAMTS-5 signaling pathway.

SOXC Transcription Factors as Diagnostic Biomarkers and Therapeutic Targets for Arthritis

“…In addition, SOX4 was elevated in inflamed arthritis patients’ synovium compared to non-inflamed synovium and then was considered to be an early diagnostic biomarker during OA pathogenesis [ 5 ]. Consistent with that, treating chondrocytes with the inflammatory agent AGEs upregulated the level of SOX4 in a dose-dependent manner in vitro [ 6 ]. Similarly, SOX4 is upregulated in OA patients and IL-1β-treated chondrocytes [ 9 ].…”

“…Mechanistically, in a dose-dependent manner, AGE-induced chondrocyte injury was found to upregulate the levels of both SOX4 and phosphorylated p38. In the meantime, treatment with the p38 inhibitor reduced AGE-induced SOX4 expression, indicating that the upregulation of SOX4 that resulted from AGE treatment is mediated by p38 [ 6 ]. However, two miRNAs were found to target SOX4, miR-31-5p and miR-373-3p.…”

“…In this regard, the classic nonsteroidal anti-inflammatory drug Feprazone was reported to have a promising effective role in treating arthritis. This has been proven mechanistically by the ability of Feprazone to reduce the degraded aggrecan in the human chondrocytes induced in TNF-α treated models, where it inhibited the loss of aggrecan via the suppression of the SOX-4/ADAMTS-5 signaling pathway [ 6 ]. In addition, we have indicated that the natural flavonoid biomolecule pinocembrin can virtually interact with SOX4 and reduces its level expression in RA mouse models leading to a reduction in arthritis symptoms [ 11 ].…”

“…SOXC is a superfamily of TFs that have been recently shown by several studies to be involved in the onset and pathogenesis of arthritis. Interestingly, the great majority of these studies have been conducted during the last 5 years [ 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 ]. The SOXC gene family encodes SOX4, SOX11, and SOX12 TFs that contain a conserved high mobility group (HMG)-box domain by which SOXC proteins are able to bind to minor grooves in the DNA.…”

The therapeutic effects of low-intensity pulsed ultrasound in musculoskeletal soft tissue injuries: Focusing on the molecular mechanism

“…IL-1β also plays a significant role in the pathogenesis of OA and is substantially expressed in OA patients (Kapoor et al, 2011). ADAMTS-5 is a catabolic enzyme that promotes EMC degradation in cartilage (Xiong et al, 2021). In addition to the combination of LIPUS with stem cells, the combination of LIPUS with other therapies can also accelerate the recovery of cartilage damage, for example, the combination with fibroblast growth factor 2 in the study of Tang et al, with low-level laser therapy in the study of Paolillo et al and with Prussian blue nanoparticles (Paolillo et al, 2018;Tang and Li, 2018;Zuo et al, 2021).…”