Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, China; Key Laboratory of Transplant Engineering and Immunology, West China Hospital, Sichuan University, Chengdu, China.
Department of Orthopedics, West China Hospital, Sichuan University, Chengdu, China.
Osteoarthritis Cartilage. 2017 Oct;25(10):1698-1707. doi: 10.1016/j.joca.2017.06.002. Epub 2017 Jun 21.
Disruptions of extracellular matrix (ECM) homeostasis are key events in the pathogenesis of osteoarthritis (OA). MicroRNA-140 (miRNA-140) is expressed specifically in cartilage and regulates ECM-degrading enzymes. Our objective in this study was to determine if intra-articular injection of miRNA-140 can attenuate OA progression in rats.
miRNA-140 levels in human normal and OA cartilage derived chondrocytes and synovial fluid were assessed by polymerase chain reaction (PCR). After primary human chondrocytes were transfected with miRNA-140 mimic or inhibitor, PCR and western blotting were performed to quantify Collagen II, MMP-13, and ADAMTS-5 expression. An OA model was induced surgically in rats, and subsequently treated with one single intra-articular injection of miRNA-140 agomir. At 4, 8, and 12 weeks after surgery, OA progression were evaluated macroscopically, histologically, and immunohistochemically in these rats.
miRNA-140 levels were significantly reduced in human OA cartilage derived chondrocytes and synovial fluid compared with normal chondrocytes and synovial fluid. Overexpressing miRNA-140 in primary human chondrocytes promoted Collagen II expression and inhibited MMP-13 and ADAMTS-5 expression. miRNA-140 levels in rat cartilage were significantly higher in the miRNA-140 agomir group than in the control group. Moreover, behavioural scores, chondrocyte numbers, cartilage thickness and Collagen II expression levels in cartilage were significantly higher, while pathological scores and MMP-13 and ADAMTS-5 expression levels were significantly lower in the miRNA-140 agomir group than in the control group.
Intra-articular injection of miRNA-140 can alleviate OA progression by modulating ECM homeostasis in rats, and may have potential as a new therapy for OA.
细胞外基质（ECM）稳态的破坏是骨关节炎（OA）发病机制中的关键事件。microRNA-140（miRNA-140）特异性表达于软骨中，并调节 ECM 降解酶。本研究旨在确定关节内注射 miRNA-140 是否可以减轻大鼠 OA 的进展。
通过聚合酶链反应（PCR）评估人正常和 OA 软骨来源软骨细胞和滑液中的 miRNA-140 水平。转染 miRNA-140 模拟物或抑制剂后，通过 PCR 和 Western blot 定量评估 Collagen II、MMP-13 和 ADAMTS-5 的表达。在大鼠中手术诱导 OA 模型，随后用 miRNA-140 激动剂进行单次关节内注射治疗。手术后 4、8 和 12 周，对这些大鼠进行大体、组织学和免疫组织化学评估 OA 进展。
与正常软骨细胞和滑液相比，人 OA 软骨来源的软骨细胞和滑液中的 miRNA-140 水平显著降低。在原代人软骨细胞中过表达 miRNA-140 可促进 Collagen II 的表达，并抑制 MMP-13 和 ADAMTS-5 的表达。miRNA-140 激动剂组大鼠软骨中的 miRNA-140 水平明显高于对照组。此外，miRNA-140 激动剂组的行为评分、软骨细胞数量、软骨厚度和 Collagen II 表达水平显著升高，而 MMP-13 和 ADAMTS-5 的表达水平显著降低。
关节内注射 miRNA-140 通过调节大鼠 ECM 稳态可减轻 OA 进展，可能作为 OA 的一种新疗法具有潜力。
