Li Ningbo, Wang Yongsheng, Wu Xuejian
Department of Orthopedics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
Department of Orthopedics, The First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, China.
Front Genet. 2022 Aug 24;13:886898. doi: 10.3389/fgene.2022.886898. eCollection 2022.
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. 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. 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. Circ_0037658 knockdown relieved IL-1β-triggered chondrocyte injury via regulating the miR-665/ADAMTS5 axis, promising an underlying therapeutic strategy for OA.
骨关节炎（OA）是一种慢性肌肉骨骼退行性疾病，给患者带来巨大痛苦，并给全球医疗资源造成巨大负担。先前的报道表明，环状RNA（circRNAs）参与了OA的发病机制。本研究的目的是探讨circ_0037658在OA细胞模型中的作用及机制。采用酶联免疫吸附测定（ELISA）检测白细胞介素-6（IL-6）和肿瘤坏死因子α（TNF-α）的含量。使用细胞计数试剂盒-8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EDU）和流式细胞术检测细胞增殖能力和凋亡情况。采用蛋白质印迹法检测Bcl-2相关X蛋白（Bax）、裂解的半胱天冬酶-3、基质金属蛋白酶13、聚集蛋白聚糖和含血小板反应蛋白基序的解聚素和金属蛋白酶（ADAMTS）5的蛋白水平。采用实时定量聚合酶链反应（RT-qPCR）检测circ_0037658、微小RNA-665（miR-665）和含血小板反应蛋白基序的解聚素和金属蛋白酶（ADAMTS）5的表达。采用双荧光素酶报告基因测定和RNA免疫沉淀（RIP）测定分析circ_0037658、miR-665和ADAMTS5之间的关系。用人软骨细胞（CHON-001细胞）用白细胞介素-1β（IL-1β）处理以建立OA细胞模型。在OA软骨样本和IL-1β处理的软骨细胞中，circ_0037658和ADAMTS5水平升高，而miR-665水平降低。此外，circ_0037658沉默促进了IL-1β处理的CHON-001细胞的增殖，并损害了炎症、凋亡和细胞外基质降解。机制上，circ_0037658作为miR-665的海绵来调节ADAMTS5的表达。circ_0037658敲低通过调节miR-665/ADAMTS5轴减轻了IL-1β引发的软骨细胞损伤，为OA提供了一种潜在的治疗策略。
