Department of Joint Orthopedics, Wujin Hospital Affiliated with Jiangsu University, Changzhou City, Jiangsu, 213002, China.
Department of Joint Orthopedics, the Wujin Clinical College of Xuzhou Medical University, Changzhou City, Jiangsu, 213002, China.
J Orthop Surg Res. 2024 Apr 15;19(1):241. doi: 10.1186/s13018-024-04694-z.
Circular RNAs (circRNAs) play an important role in osteoarthritis (OA). However, the role of circRNA in OA is still unclear. Here, we explored the role and mechanism of circ_0044235 in OA.
CHON-001 cells were treated with IL-1β to establish OA model in vitro. The levels of circ_0044235, miR-375 and phosphoinositide 3-kinase (PI3K) regulatory subunit 3 (PIK3R3) were detected by quantitative real-time PCR. Cell count kit-8 assay and flow cytometry assay were used to detect cell viability and apoptosis. The concentrations of inflammation factors were determined by enzyme-linked immunosorbent assay. Western blot was used to detect protein levels. The interaction between miR-375 and circ_0044235 or PIK3R3 was analyzed by dual-luciferase reporter assay and RNA immunoprecipitation assay.
Circ_0044235 was significantly decreased in OA cartilage tissue and IL-1β-treated CHON-001 cells. Overexpression of circ_0044235 promoted IL-1β-stimulated CHON-001 cell viability and inhibited apoptosis, inflammation, and extracellular matrix (ECM) degradation. In mechanism analysis, circ_0044235 could act as a sponge for miR-375 and positively regulate PIK3R3 expression. In addition, miR-375 ameliorated the effect of circ_0044235 overexpression on IL-1β-mediated CHON-001 cells injury. In addition, miR-375 inhibition mitigated IL-1β-induced CHON-001 cell injury, while PIK3R3 silencing restored the effect.
Circ_0044235 knockdown alleviated IL-1β-induced chondrocytes injury by regulating miR-375/PIK3R3 axis, confirming that circ_0044235 might be a potential target for OA treatment.
环状 RNA（circRNA）在骨关节炎（OA）中发挥重要作用。然而，circRNA 在 OA 中的作用尚不清楚。在这里，我们探讨了 circ_0044235 在 OA 中的作用和机制。
用白细胞介素-1β（IL-1β）处理 CHON-001 细胞在体外建立 OA 模型。用实时定量 PCR 检测 circ_0044235、miR-375 和磷脂酰肌醇 3-激酶（PI3K）调节亚基 3（PIK3R3）的水平。用细胞计数试剂盒-8 检测和流式细胞术检测细胞活力和凋亡。用酶联免疫吸附试验测定炎症因子浓度。用 Western blot 检测蛋白水平。用双荧光素酶报告基因检测和 RNA 免疫沉淀检测分析 miR-375 与 circ_0044235 或 PIK3R3 的相互作用。
OA 软骨组织和 IL-1β 处理的 CHON-001 细胞中 circ_0044235 显著降低。circ_0044235 的过表达促进了 IL-1β 刺激的 CHON-001 细胞活力，并抑制了凋亡、炎症和细胞外基质（ECM）降解。在机制分析中，circ_0044235 可以作为 miR-375 的海绵，并正向调节 PIK3R3 的表达。此外，miR-375 改善了 circ_0044235 过表达对 IL-1β 介导的 CHON-001 细胞损伤的作用。此外，miR-375 抑制减轻了 IL-1β 诱导的 CHON-001 细胞损伤，而 PIK3R3 沉默恢复了这种作用。
circ_0044235 的敲低通过调节 miR-375/PIK3R3 轴缓解了 IL-1β 诱导的软骨细胞损伤，证实 circ_0044235 可能是 OA 治疗的潜在靶点。
