Department of Orthopedics, the Third Affiliated Hospital (the Affiliated Luohu Hospital) of Shenzhen University, Shenzhen, Guangdong, 518001, People's Republic of China.
The Spine Department, Orthopaedic Center, Guangdong Second Provincial General Hospital, No. 466, Mid Xingang Road, Haizhu District, Guangzhou, Guangdong, 510317, People's Republic of China.
Cell Biol Toxicol. 2023 Jun;39(3):1099-1118. doi: 10.1007/s10565-021-09665-2. Epub 2021 Oct 25.
To probe the performance of miR-337-3p on the facet joint osteoarthritis (FJOA) and its underlying mechanism.
qRT-PCR and Western blot were utilized to analyze the levels of miR-337-3p and DUSP1 in the synovial tissues from 36 FJOA patients and 10 healthy controls. The human synovial fibroblasts of FJOA were isolated and cultured followed by cell transfection. Then, cells were exposed to 10 ng/mL of IL-1β to induce inflammatory response of synovial fibroblasts. The alternation on cell biological function in cell models was determined. The binding of miR-337-3p and SKP2 was predicted by StarBase, TargetScan, DIANA-microT and miRmap, and further verified by RIP assay and dual-luciferase reporter assay. Co-IP experiment and ubiquitination assay were used to display the binding of SKP2 and DUSP1 as well as the ubiquitination and degradation of DUSP1. After that, the FJOA rat model was established and miR-337-3p mimic or negative control was given to rats by tail vein injection. The pathological changes of synovial tissues, synovitis score, and inflammation level in rats were assessed.
The low expressions of miR-337-3p and DUSP1 were noticed in the synovial tissues of FJOA patients and in IL-1β-induced synovial fibroblasts, and highly expressed p-p38 MAPK was noticed. Upregulation of miR-337-3p/DUSP1 or downregulation of SKP2 inhibited IL-1β-induced proliferation and inflammatory response of synovial fibroblasts. SKP2 was the target gene of miR-337-3p, and SKP2 induced the ubiquitination and degradation of DUSP1. MiR-337-3p exerted a protective effect on FJOA rats by alleviating damage of rat synovial tissues, promoting cell apoptosis and repressing inflammatory response.
MiR-337-3p plays a protective role in FJOA by negatively targeting SKP2 to suppress DUSP1 ubiquitination and inactivate the p38 MAPK pathway.
探讨 miR-337-3p 在小关节骨关节炎（FJOA）中的作用及其机制。
利用 qRT-PCR 和 Western blot 检测 36 例 FJOA 患者和 10 例健康对照的滑膜组织中 miR-337-3p 和 DUSP1 的水平。分离并培养人 FJOA 滑膜成纤维细胞，然后进行细胞转染。接着，用 10ng/ml 的 IL-1β 诱导滑膜成纤维细胞的炎症反应。检测细胞模型中细胞生物学功能的改变。通过 StarBase、TargetScan、DIANA-microT 和 miRmap 预测 miR-337-3p 和 SKP2 的结合，并用 RIP 实验和双荧光素酶报告基因实验进一步验证。用 Co-IP 实验和泛素化实验显示 SKP2 和 DUSP1 的结合以及 DUSP1 的泛素化和降解。然后，建立 FJOA 大鼠模型，通过尾静脉注射 miR-337-3p 模拟物或阴性对照，评估大鼠滑膜组织的病理变化、滑膜炎评分和炎症水平。
在 FJOA 患者的滑膜组织和 IL-1β 诱导的滑膜成纤维细胞中，miR-337-3p 和 DUSP1 的表达水平较低，而 p-p38 MAPK 的表达水平较高。上调 miR-337-3p/DUSP1 或下调 SKP2 抑制了 IL-1β 诱导的滑膜成纤维细胞的增殖和炎症反应。SKP2 是 miR-337-3p 的靶基因，SKP2 诱导 DUSP1 的泛素化和降解。miR-337-3p 通过减轻大鼠滑膜组织损伤、促进细胞凋亡和抑制炎症反应，对 FJOA 大鼠发挥保护作用。
miR-337-3p 通过负向靶向 SKP2 抑制 DUSP1 泛素化和失活 p38 MAPK 通路，在 FJOA 中发挥保护作用。
