AIMS The study aimed to investigate the roles of miR-483-5p and IGF2 in osteoclast formation. METHODS Blood and bone tissues were collected from osteoporosis and non-osteoporosis patients with hip fractures for gene expression analysis. CD14 + peripheral blood mononuclear cells (PBMCs) were isolated for differentiating osteoclasts. MiR-483-5p mimic and inhibitor was transfected into CD14 + PBMCs, respectively. Predicted by TargetScan and verified by Dual-luciferase reporter assay system, insulin-like growth factor-2 (IGF2) could be targeted by miR-483-5p. IGF2 expression vector was co-transfected with miR-483-5p mimic to study the role of IGF2 in miR-483-5p affecting osteoclast differentiation. Flow cytometry was performed for cell apoptosis analysis. RESULTS High-expressed miR-483-5p and low-expressed IGF2 were frequently found in the serums and bone tissues derived from osteoporotic patients. We found that up-regulation of miR-483-5p in CD14 + PBMCs notably increased the number of TRAP-positive cells, at the same time, the expression levels of TRAP, nuclear factor of activated T-cells (NFATc1), cytoplasmic 1 (NFAT2) and Cathepsin K (CTSK) were also up-regulated. However, overexpressed IGF2 effectively reversed such effects produced by up-regulation of miR-483-5p on osteoclastogenesis-related factors in CD14 + PBMCs. Moreover, forced expression of IGF2 could also enhance apoptosis of osteoclasts reduced by miR-483-5p. CONCLUSIONS Our study suggests that miRNA-483-5p is involved in the pathogenesis of osteoporosis by promoting osteoclast differentiation.

中文翻译：

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MiRNA-483-5p通过促进破骨细胞分化而参与骨质疏松症的发病机制。

目的本研究旨在研究miR-483-5p和IGF2在破骨细胞形成中的作用。方法收集骨质疏松症和非骨质疏松症髋部骨折患者的血液和骨组织，进行基因表达分析。分离CD14 +外周血单个核细胞（PBMC）以分化破骨细胞。将MiR-483-5p模拟物和抑制剂分别转染到CD14 + PBMC中。由TargetScan预测并由双荧光素酶报告基因检测系统验证，miR-483-5p可以靶向胰岛素样生长因子2（IGF2）。将IGF2表达载体与miR-483-5p模拟物共转染，以研究IGF2在miR-483-5p中影响破骨细胞分化的作用。进行流式细胞术以进行细胞凋亡分析。结果在骨质疏松症患者的血清和骨组织中经常发现高表达的miR-483-5p和低表达的IGF2。我们发现，CD14 + PBMC中miR-483-5p的上调显着增加了TRAP阳性细胞的数量，同时，TRAP的表达水平，活化T细胞的核因子（NFATc1），细胞质1 （NFAT2）和组织蛋白酶K（CTSK）也被上调。然而，过表达的IGF2有效逆转了miR-483-5p上调对CD14 + PBMC中破骨细胞生成相关因子的影响。此外，IGF2的强制表达还可以增强被miR-483-5p减少的破骨细胞凋亡。结论我们的研究表明，miRNA-483-5p通过促进破骨细胞分化而参与骨质疏松的发病机制。