Circ_0022383 alleviates IL-1β-induced apoptosis, inflammation and extracellular matrix degeneration in osteoarthritis cell model by miR-3619-5p/SIRT1 axis.

Circular RNAs (circRNAs) have been identified to play roles in cartilage homeostasis and chondrocyte development, and be associated with osteoarthritis (OA) pathophysiology. Here, we aimed to investigate the role and mechanism of circ_0022383 on OA progression. Chondrocytes in functional groups were treated with interleukin (IL)-1β. The levels of genes and proteins were assayed by qRT-PCR and western blotting. Cell proliferation and apoptosis were evaluated by cell counting kit-8 assay, 5-Ethynyl-2'-deoxyuridine (Edu) assay, and flow cytometry, respectively. The inflammation and extracellular matrix (ECM) degeneration were determined by assessing the activity of IL-6, tumor necrosis factor (TNF-α), Aggrecan (ACAN), collagen type II α 1 chain (COL2A1) and ADAMTS5 proteins. The binding between miR-3619-5p and circ_0022383 or silent information regulator 1 (SIRT1) was confirmed by dual-luciferase reporter, RIP and RNA pull-down assays. Circ_0022383 expression was lower in the cartilages of OA patients and IL-1β-induced primary chondrocytes. Functionally, ectopic overexpression of circ_0022383 alleviated IL-1β-induced proliferation arrest, apoptosis, the release of IL-6 and TNF-α, as well as the decrease of ACAN and COL2A1 and the increase of ADAMTS5 level in chondrocytes. Mechanistically, circ_0022383 acted as a sponge for miR-3619-5p, which was verified to target SIRT1. Rescue experiments showed that miR-3619-5p up-regulation reversed the protective effects of circ_0022383 on IL-1β-stimulated chondrocytes. Additionally, miR-3619-5p inhibition abolished IL-1β-induced apoptosis, inflammation and ECM degeneration in chondrocytes, which were counteracted by SIRT1 silencing. Circ_0022383 protected chondrocytes from IL-1β-induced apoptosis, inflammation and ECM degeneration by miR-3619-5p/SIRT1 axis, inspiring future therapy development for OA prevention.

Qian L ，Yu B ，Chen T ，Chen K ，Ma Z ，Wang Y ，Sun B ... -  《-》

Circ_0001103 alleviates IL-1β-induced chondrocyte cell injuries by upregulating SIRT1 via targeting miR-375.

Osteoarthritis (OA) is a common inflammatory disease characterized by articular cartilage degeneration and injury. Circular RNAs (circRNAs) are widely involved in the development of human diseases, including OA. The objective of this study was to investigate the function and functional mechanism of circ_0001103 in OA. Cell model of OA was established by treating chondrocytes with interleukin-1β (IL-1β). The expression of circ_0001103, miR-375 and sirtuin 1 (SIRT1) mRNA was measured using quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability was assessed using cell counting kit-8 (CCK-8) assay. Cell apoptosis was determined using flow cytometry assay. The expression levels of inflammatory factors were quantified by qRT-PCR. The expression of extracellular matrix (ECM) metabolism-related markers, including Collagen Type II Alpha 1 Chain (COL2A1) and A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4), was detected by western blot. Predicted target relationship between miR-375 and circ_0001103 or SIRT1 by the bioinformatics tools was validated by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. Circ_0001103 was downregulated in OA tissues and IL-1β-induced chondrocytes. Overexpression of circ_0001103 attenuated IL-1β-induced chondrocyte apoptosis, inflammatory responses and ECM degradation. MiR-375 was targeted by circ_0001103, and miR-375 could bind to SIRT1. Circ_0001103 overexpression increased the expression of SIRT1 by suppressing miR-375. Rescue experiments suggested that miR-375 restoration reversed the effects of circ_0001103 overexpression, and SIRT1 knockdown overturned the effects of miR-375 inhibition. Circ_0001103 governed the miR-375/SIRT1 axis to ameliorate IL-1β-induced chondrocyte injuries, implying that circ_0001103 was a promising biomarker in OA pathogenesis.

Zhang M ，Mou L ，Liu S ，Sun F ，Gong M ... -  《-》

CircRNA circ_SEC24A upregulates DNMT3A expression by sponging miR-26b-5p to aggravate osteoarthritis progression.

Osteoarthritis (OA) is a chronic degenerative disease characterized by degeneration and injury of articular cartilage. Circular RNA_SEC24A (circ_SEC24A; circBase ID: hsa_circ_0005105) is upregulated and promotes multiple tumor processes. However, its role in OA progression remained mostly unknown. Quantitative real-time PCR (qRT-PCR) was used to detect the RNA expression of circ_SEC24A, miR-26b-5p and DNA methyltransferase 3 alpha (DNMT3A). Cell proliferation was verified by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) and 5-ethynyl-2'-deoxyuridine (EdU) assays. Flow cytometry was used to detect apoptosis. Western blot was used to detect protein expression of DNMT3A, proliferating cell nuclear antigen (PCNA), extracellular matrix (ECM) proteins (Collagen II and Aggrecan), and ECM degrading enzymes (matrix metalloproteinase-13 [MMP13] and metallopeptidase with thrombospondin type 1 motif 5 [ADAMTS5]). The target relationship between miR-26b-5p and circ_SEC24A or DNMT3A was predicted by Statbase3.0 or TargetScan and confirmed by dual-luciferase reporter assay, RNA pull-down assay and RNA immunoprecipitation. Circ_SEC24A was upregulated in osteoarthritic cartilage tissues and IL-1β-induced chondrocytes, accompanying with miR-26b-5p downregulation and DNMT3A upregulation. Circ_SEC24A expression was resistant to RNase R digestion and mainly expressed in the cytoplasm. Interfering circ_SEC24A abolished IL-1β-induced effects on proliferation inhibition, apoptosis, and ECM degradation in chondrocytes, but overexpressing circ_SEC24A had the opposite effects. Inhibiting miR-26b-5p counteracted but upregulating miR-26a-5p mimicked the functions of circ_SEC24A silencing. Reinforcing DNMT3A reversed miR-26b-5p overexpression's role in IL-1β-induced chondrocytes. Mechanically, circ_SEC24A and DNMT3A were competitive endogenous RNAs (ceRNAs) for miR-26b-5p. Circ_SEC24A was a promoting factor for IL-1β-induced OA progression via circ_SEC24A/miR-26b-5p/DNMT3A ceRNA axis.

Zhang Z ，Yang B ，Zhou S ，Wu J ... -  《-》

Knockdown of circ-PRKCH alleviates IL-1β-treated chondrocyte cell phenotypic changes through modulating miR-502-5p/ADAMTS5 axis.

Liu Z ，Cao J ，Zhang L ，Li J ，Yan T ，Zhou P ，Zhang S ... -  《-》

Circ_0134111 knockdown relieves IL-1β-induced apoptosis, inflammation and extracellular matrix degradation in human chondrocytes through the circ_0134111-miR-515-5p-SOCS1 network.

Osteoarthritis (OA) is characterized by chondrocyte injury and dysfunction, such as excessive apoptosis, inflammatory response and extracellular matrix (ECM) degradation. Circular RNA (circRNA) deregulation is reported to be involved in OA. Our study aimed to explore the role of circ_0134111 in OA. Human chondrocytes were treated with interleukin-1β (IL-1β) to mimic OA cell model. The expression of circ_0134111, miR-515-5p and suppressor of cytokine signaling 1 (SOCS1) mRNA was measured by real-time quantitative polymerase chain reaction (RT-qPCR), and the protein levels of SOCS1 and apoptosis-/inflammation-/ECM-related markers were determined by western blot. Cell proliferation and cell apoptosis were assessed using cell counting kit-8 (CCK-8) and flow cytometry assay, respectively. For mechanism analysis, the predicted interaction between miR-515-5p and circ_0134111 or SOCS1 was verified by dual-luciferase reporter assay, pull-down assay and RNA immunoprecipitation (RIP) assay. Rescue experiments were performed to explore the interplay between miR-515-5p and circ_0134111 or SOCS1. Circ_0134111 was overexpressed in OA cartilage tissues and IL-1β-induced chondrocytes. IL-1β-induced chondrocyte apoptosis, inflammatory responses and ECM degradation were alleviated by circ_0134111 knockdown or miR-515-5p restoration. Circ_0134111 acted as miR-515-5p sponge to regulate miR-515-5p expression, and miR-515-5p deficiency reversed the effects of circ_0134111 knockdown in IL-1β-induced chondrocytes. MiR-515-5p directly bound to SOCS1, and circ_0134111 decoyed miR-515-5p to increase SOCS1 level. MiR-515-5p restoration alleviated IL-1β-induced chondrocyte apoptosis, inflammatory responses and ECM degradation, While SOCS1 overexpression partly abolished these effects. Circ_0134111 knockdown alleviated apoptosis, inflammatory responses and ECM degradation in OA cell model by mediating the miR-515-5p-SOCS1 network, hinting that circ_0134111 was involved in OA progression.

Wu R ，Zhang F ，Cai Y ，Long Z ，Duan Z ，Wu D ，Zhou Y ，Wang Q ... -  《-》