Ou Liang, Huang Weichen, Zhang Tiantian, Xu Daoqing, Kong Dezhong, Meng Yingfu
Hunan Academy of Chinese Medicine, Changsha, 410006, China; Department of Orthopedic, The Second Affiliated Hospital of Guizhou University of Chinese Medicine, Guiyang, Guizhou Province 550003, China.
Department of Orthopedic, The Second Affiliated Hospital of Guizhou University of Chinese Medicine, Guiyang, Guizhou Province 550003, China.
Transpl Immunol. 2023 Apr;77:101747. doi: 10.1016/j.trim.2022.101747. Epub 2022 Nov 25.
Osteoarthritis (OA) was a chronic degenerative joint disease. The dysregulation of circular RNAs (circRNAs) has been identified in OA progression. However, the function and regulation mechanism of circ_0114876 in OA remains largely unknown.
Firstly, we used LPS-treated C28/I2 cells as a cellular model of OA. Quantificational real-time polymerase chain reaction (qRT-PCR) was used to determine the expression levels of circ_0114876, miRNA-1227-3p, and ADAM10 in OA chondrocytes. Cell Counting Kit-8 (CCK8), 5-ethynyl-20-deoxyuridine (EdU) incorporation assays, flow cytometry, Enzyme-linked immunosorbent assay (ELISA) kit, and western blot were applied to confirm cell proliferation, apoptosis, inflammation, and extracellular matrix. of circ_0114876 in vitro. The interaction between circ_0114876 and its downstream target (miR-1227-3p) and mRNA target ADAM metallopeptidase domain 10 (ADAM10), was evaluated by luciferase assay and RNA immunoprecipitation (RIP) assay.
Circ_0114876 and ADAM10 were upregulated and miR-1227-3p was decreased in OA tissues and LPS-treated chondrocytes. Low expression of circ_0114876 promoted proliferation and inhibited apoptosis, inflammation, and extracellular matrix of the LPS-treated chondrocytes. Mechanistically, circ_0114876 functioned in human chondrocytes through targeting miR-1227-3p and ADAM10. Furthermore, miRNA-1227-3p inhibitor reversed the effect of circ_0114876 knockdown on the OA chondrocytes, and ADAM10 overexpression reversed the effect of miR-1227-3p mimic on the OA chondrocytes.
Circ_0114876 was increased in OA tissues and cells. Circ_0114876 facilitated the progression in the LPS-induced OA cell model via regulating the miR-1227-3p/ADAM10 axis. This study would provide a potentially effective therapeutic strategy for OA progression.
骨关节炎（OA）是一种慢性退行性关节疾病。环状RNA（circRNAs）的失调已在OA进展过程中被发现。然而，circ_0114876在OA中的功能和调控机制仍 largely未知。
首先，我们使用脂多糖（LPS）处理的C28/I2细胞作为OA的细胞模型。采用定量实时聚合酶链反应（qRT-PCR）来测定OA软骨细胞中circ_0114876、miRNA-1227-3p和ADAM10的表达水平。应用细胞计数试剂盒-8（CCK8）、5-乙炔基-2'-脱氧尿苷（EdU）掺入试验、流式细胞术、酶联免疫吸附测定（ELISA）试剂盒和蛋白质免疫印迹法来体外确认circ_0114876对细胞增殖、凋亡、炎症和细胞外基质的影响。通过荧光素酶报告基因试验和RNA免疫沉淀（RIP）试验评估circ_0114876与其下游靶点（miR-1227-3p）和mRNA靶点ADAM金属蛋白酶结构域10（ADAM10）之间的相互作用。
在OA组织和LPS处理的软骨细胞中，circ_0114876和ADAM10上调，而miR-1227-3p下调。circ_0114876的低表达促进了LPS处理的软骨细胞的增殖，并抑制了其凋亡、炎症和细胞外基质。机制上，circ_0114876通过靶向miR-1227-3p和ADAM10在人软骨细胞中发挥作用。此外，miRNA-1227-3p抑制剂逆转了circ_0114876敲低对OA软骨细胞的影响，而ADAM10过表达逆转了miR-1227-3p模拟物对OA软骨细胞的影响。
circ_0114876在OA组织和细胞中表达增加。circ_0114876通过调节miR-1227-3p/ADAM10轴促进LPS诱导的OA细胞模型的进展。本研究将为OA进展提供一种潜在有效的治疗策略。
