miRNA-483-5p 通过促进破骨细胞分化参与骨质疏松症的发病机制。

MiRNA-483-5p is involved in the pathogenesis of osteoporosis by promoting osteoclast differentiation.

机构信息

Department of Orthopedics, Suizhou Hospital, Hubei University of Medicine, China.

出版信息

Mol Cell Probes. 2020 Feb;49:101479. doi: 10.1016/j.mcp.2019.101479. Epub 2019 Nov 6.

DOI:10.1016/j.mcp.2019.101479

PMID:31706013

Abstract

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.

摘要

目的

本研究旨在探讨 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+外周血单核细胞（PBMC）进行破骨细胞分化。分别转染 miR-483-5p 模拟物和抑制剂至 CD14+PBMC。通过 TargetScan 预测和双荧光素酶报告基因检测系统验证，胰岛素样生长因子 2（IGF2）可被 miR-483-5p 靶向。将 IGF2 表达载体与 miR-483-5p 模拟物共转染，研究 IGF2 在 miR-483-5p 影响破骨细胞分化中的作用。流式细胞术进行细胞凋亡分析。

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 的上调显著增加了 TRAP 阳性细胞的数量，同时，TRAP、活化 T 细胞核因子（NFATc1）、细胞质 1（NFAT2）和组织蛋白酶 K（CTSK）的表达水平也上调。然而，过表达 IGF2 可有效逆转 miR-483-5p 对 CD14+PBMC 中破骨细胞相关因子的上调作用。此外，IGF2 的强制表达也可增强 miR-483-5p 降低的破骨细胞凋亡。