Department of Orthopaedics, Ningbo First Hospital.
Department of Orthopaedics, The First Affiliated Hospital of Soochow University.
Biol Pharm Bull. 2020 May 1;43(5):810-816. doi: 10.1248/bpb.b19-00959. Epub 2020 Feb 26.
Osteoarthritis is a common disease character with progressive destruction of cartilage. MicroRNA (miR)-140-3p was validated as a biomarker for osteoarthritis. However, the mechanism by which miRNA-140-3p regulates osteoarthritis remains unclear. Thus, this study aims to evaluate the potential function of miRNA-140-3p during the pathogenesis of osteoarthritis. MiRNA-140-3p expression in tissue and CHON-001 chondrocyte cells was determined with quantitative real time (qRT)-PCR. In vitro osteoarthritis model was established by treatment of the chondrocyte cells CHON-001 with interleukin (IL)-1β. Cell proliferation and apoptosis were measured with cell counting kit-8 (CCK8) and Annexin V/propidium iodide (PI) apoptosis assay, respectively. Protein expressions were evaluated using Western blot. The target gene of miR-140-3p was predicted using Targetscan and miRDB. MiR-140-3p was downregulated in knee tissue from patients with osteoarthritis. IL-1β inhibited the proliferation of CHON-001 cells via inducing apoptosis. In addition, IL-1β significantly inhibited the expressions of collagen II and aggrecan and increased the level of matrix metalloproteinase (MMP)13. However, the effects of IL-1β could be ameliorated by the addition of miR-140-3p mimics. Moreover, luciferase reporter assay demonstrated CXCR4 as a target gene of miR-140-3p. IL-1β-induced upregulation of CXCR4 could be blocked by miR-140-3p mimics. Our study indicated that miR-140-3p could suppress the progression of osteoarthritis by directly targeting CXCR4. Therefore, miR-140-3p might serve as a potential therapeutic target for the treatment of osteoarthritis.
骨关节炎是一种以软骨进行性破坏为特征的常见疾病。微小 RNA（miR）-140-3p 已被验证为骨关节炎的生物标志物。然而，miRNA-140-3p 调节骨关节炎的机制尚不清楚。因此，本研究旨在评估 miRNA-140-3p 在骨关节炎发病机制中的潜在功能。采用实时定量聚合酶链反应（qRT-PCR）检测组织和 CHON-001 软骨细胞中 miR-140-3p 的表达。用白细胞介素（IL）-1β处理软骨细胞 CHON-001 建立体外骨关节炎模型。采用细胞计数试剂盒-8（CCK8）和 Annexin V/碘化丙啶（PI）凋亡检测法分别检测细胞增殖和凋亡。采用 Western blot 法检测蛋白表达。采用 Targetscan 和 miRDB 预测 miR-140-3p 的靶基因。骨关节炎患者膝关节组织中 miR-140-3p 表达下调。IL-1β 通过诱导细胞凋亡抑制 CHON-001 细胞的增殖。此外，IL-1β 显著抑制胶原 II 和聚集蛋白聚糖的表达，增加基质金属蛋白酶（MMP）13 的水平。然而，添加 miR-140-3p 模拟物可以改善 IL-1β 的作用。此外，荧光素酶报告基因检测表明 CXCR4 是 miR-140-3p 的靶基因。miR-140-3p 模拟物可阻断 IL-1β 诱导的 CXCR4 上调。本研究表明，miR-140-3p 可通过直接靶向 CXCR4 抑制骨关节炎的进展。因此，miR-140-3p 可能成为治疗骨关节炎的潜在治疗靶点。
