Knockdown of Circ_0037658 Alleviates IL-1β-Induced Osteoarthritis Progression by Serving as a Sponge of miR-665 to Regulate ADAMTS5

Abstract

Background: Osteoarthritis (OA) is a chronic musculoskeletal degeneration disease which brings great pain to patients and a tremendous burden on the world’s medical resources. Previous reports have indicated that circular RNAs (circRNAs) are involved in the pathogenesis of OA. The purpose of this study was to explore the role and mechanism of circ_0037658 in the OA cell model.Methods: The content of interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) was measured using enzyme-linked immunosorbent assay (ELISA). Cell proliferation ability and apoptosis were detected using Cell Counting Kit-8 (CCK-8), 5-ethynyl-2′-deoxyuridine (EDU), and flow cytometry assays. Western blot assay was used to measure the protein levels of Bcl-2-related X protein (Bax), cleaved-caspase-3, MMP13, Aggrecan, and ADAMTS5. The expression of circ_0037658, microRNA-665 (miR-665), and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 5 was detected using real-time quantitative polymerase chain reaction (RT-qPCR). Dual-luciferase reporter assay and RNA Immunoprecipitation (RIP) assay were manipulated to analyze the relationships of circ_0037658, miR-665, and ADAMTS5.Results: Human chondrocytes (CHON-001 cells) were treated with interleukin-1β (IL-1β) to establish an OA cell model. Circ_0037658 and ADAMTS5 levels were increased, and miR-665 was decreased in OA cartilage samples and IL-1β-treated chondrocyte cells. Moreover, circ_0037658 silencing promoted proliferation and impaired inflammation, apoptosis, and ECM degradation in IL-1β-treated CHON-001 cells. Mechanically, circ_0037658 acted as a sponge for miR-665 to regulate ADAMTS5 expression.Conclusion: Circ_0037658 knockdown relieved IL-1β-triggered chondrocyte injury via regulating the miR-665/ADAMTS5 axis, promising an underlying therapeutic strategy for OA.

ADAMTS5 Promotes Permeability of the Blood-Brain Barrier during Treponema pallidum Subspecies pallidum Invading the Central Nervous System

“…Given the documented ability of IL-1β to induce ADAMTS5 up-regulation in human chondrocytes, , we determined the expression level of IL-1β in Tp-treated hCMEC/D3 cells. After stimulation with Tp at an MOI of 50:1 for 24 h, the expression of IL-1β was detected by RT-qPCR and ELISA.…”

“…Considering that IL-1β had been demonstrated to induce the up-regulation of ADAMTS5 in human chondrocytes, , we also investigated whether Tp regulated the expression of ADAMTS5 through IL-1β. Our findings revealed that Tp induced the production of IL-1β in hCMEC/D3 cells and that recombinant human IL-1β promoted the expression of ADAMTS5 in these cells, which was consistent with previous studies. , Moreover, the inhibition of IL-1β using diacerein significantly reduced the production of ADAMTS5. These data suggested that Tp enhanced ADAMTS5 expression by inducing the production of IL-1β, and IL-1β could regulate the expression of ADAMTS5, which might indirectly affect the subsequent effects of ADAMTS5.…”

“…ADAMTS5, a secreted metalloprotease, was known for its capacity to degrade aggrecan . Since its discovery in 1999, extensive research had focused primarily on conditions such as osteoarthritis and joint degeneration. − Notably, it had been observed that interleukin-1β (IL-1β) could up-regulate the expression of ADAMTS5 in human chondrocytes. , A deeper understanding of ADAMTS5 revealed its involvement in the degradation of a critical structural component of the vascular endothelium’s extracellular matrix, known as the endothelial glycocalyx . The endothelial glycocalyx was an extracellular matrix layer that covered the surface of endothelial cells, primarily composed of heparan sulfate, chondroitin sulfate, and hyaluronic acid .…”

Bioinformatics and Integrative Experimental Method to Identifying and Validating Co-Expressed Ferroptosis-Related Genes in OA Articular Cartilage and Synovium

“…It was shown that miR-665 expression is decreased in the OA chondrocyte model, while overexpression of miR-665 significantly increased chondrocyte proliferation and alleviated chondrocyte inflammation, apoptosis and ECM degradation. 59 The Wnt signaling pathway influences the progression of OA, and miR-665 modulates cell proliferation and apoptosis through its impact on the Wnt/β-Catenin signaling pathway. 60,61 Blocking miR-1972 expression attenuated the production of synovial invasive vascular opacities in RA.…”

Circ_0044235 regulates the development of osteoarthritis by the modulation of miR-375/PIK3R3 axis

“…Cells were cultured in DMEM medium (Gibco, Carlsbad, CA, USA) containing 10% FBS (Gibco) at 37℃ with 5% CO 2 . In this, 10 ng/mL IL-1β was used to stimulate CHON-001 cells for 24 h to construct OA cell models [ 24 , 25 ].…”