The Second Department of Orthopaedics, CangZhou Central Hospital, Cangzhou, China.
Eur Rev Med Pharmacol Sci. 2020 Apr;24(8):4095-4102. doi: 10.26355/eurrev_202004_20987.
To illustrate the role of micro ribonucleic acid (miR)-330-5p in regulating osteogenesis through biglycan (Bgn)-mediated bone morphogenetic protein (BMP)/Smad pathway.
A mouse model of osteoporosis (OP) was established by ovariectomy (OVX). BMD and miR-330-5p levels in mice undergoing sham operation or OVX were determined. BMD and BV/TV in OP mice with in vivo knockdown of miR-330-5p were measured by Micro-CT. After silencing of miR-330-5p in mouse primary bone marrow stromal cells (BMSCs), expression changes in osteogenesis-associated genes, ALP activity, and mineralization ability were assessed. Subsequently, the interaction between miR-330-5p and Bgn was examined by Dual-Luciferase reporter gene assay and Western blotting. Then, Bgn levels in BMSCs undergoing osteogenesis at different time points were measured. At last, the regulatory effects of miR-330-5p/Bgn axis on the BMP/Smad pathway, ALP activity, and mineralization ability in BMSCs were evaluated.
BMD was decreased and miR-330-5p was upregulated in OP mice. OP mice with in vivo knockdown of miNA-330-5p presented higher BMD and BV/TV than controls. Transfection with miR-330-5p inhibitor upregulated osteogenesis-associated genes, ALP activity, and mineralization ability in BMSCs. Bgn was time-dependently upregulated in BMSCs undergoing osteogenesis, which was indicated to be the target gene of miR-330-5p. Besides, Bgn level was negatively regulated by miR-330-5p. Importantly, Bgn was able to reverse the regulatory effects of miR-330-5p on the BMP/Smad pathway, ALP activity, and mineralization ability in BMSCs.
Knockdown of miR-330-5p facilitates osteogenesis in BMSCs through the Bgn-induced BMP/Smad pathway, thus alleviating the progression of OP.
通过核心蛋白聚糖（Bgn）介导的骨形态发生蛋白（BMP）/Smad 通路阐明微小 RNA（miR）-330-5p 在调控成骨中的作用。
通过卵巢切除术（OVX）建立骨质疏松症（OP）小鼠模型。检测假手术或 OVX 小鼠的骨密度（BMD）和 miR-330-5p 水平。通过 Micro-CT 测量体内敲低 miR-330-5p 的 OP 小鼠的 BMD 和骨体积/骨小梁体积（BV/TV）。沉默小鼠原代骨髓基质细胞（BMSCs）中的 miR-330-5p 后，评估成骨相关基因的表达变化、碱性磷酸酶（ALP）活性和矿化能力。然后，通过双荧光素酶报告基因检测和 Western blot 检测 miR-330-5p 与 Bgn 之间的相互作用。接着，测量不同时间点成骨过程中 BMSCs 中的 Bgn 水平。最后，评估 miR-330-5p/Bgn 轴对 BMSCs 中 BMP/Smad 通路、ALP 活性和矿化能力的调节作用。
OP 小鼠的 BMD 降低，miR-330-5p 上调。体内敲低 miR-330-5p 的 OP 小鼠的 BMD 和 BV/TV 高于对照组。miR-330-5p 抑制剂转染上调了 BMSCs 中的成骨相关基因、ALP 活性和矿化能力。Bgn 在成骨的 BMSCs 中呈时间依赖性上调，提示其为 miR-330-5p 的靶基因。此外，Bgn 水平受 miR-330-5p 负调控。重要的是，Bgn 能够逆转 miR-330-5p 对 BMSCs 中 BMP/Smad 通路、ALP 活性和矿化能力的调节作用。
敲低 miR-330-5p 通过 Bgn 诱导的 BMP/Smad 通路促进 BMSCs 成骨，从而缓解 OP 的进展。
