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miR-140-3p 通过靶向 CXCR4 改善骨关节炎的进展。

MiR-140-3p Ameliorates the Progression of Osteoarthritis via Targeting CXCR4.

机构信息

Department of Orthopaedics, Ningbo First Hospital.

Department of Orthopaedics, The First Affiliated Hospital of Soochow University.

出版信息

Biol Pharm Bull. 2020 May 1;43(5):810-816. doi: 10.1248/bpb.b19-00959. Epub 2020 Feb 26.

Abstract

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.

摘要

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。

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