Department of Orthopedic, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, Hubei, China.
Department of Cardiovascular, Wuhan Hospital of Traditional Chinese Medicine, Wuhan, Hubei, China.
Bioengineered. 2021 Dec;12(2):11756-11767. doi: 10.1080/21655979.2021.2009618.
MicroRNAs (miRNAs) regulate osteogenic differentiation and influence osteoporosis (OP). The aim of this study was to determine the potential role of miR-874-3p in OP. The expression levels of miR-874-3p and leptin (LEP) in the femoral neck trabeculae of 35 patients with or without OP were measured by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The effects of miR-874-3p or LEP on the cell proliferation and alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), and osterix (OSX) levels were observed by upregulating miR-874-3p in human bone marrow mesenchymal stem cells (hBMSCs). Additionally, calcium deposition levels were evaluated using alizarin red staining (ARS). Molecular mechanisms of miR-874-3p and LEP underlying the osteogenic differentiation of hBMSCs were also evaluated using bioinformatics analysis, luciferase reporter assays, and RNA pull-down assays. The miR-874-3p levels were significantly lower in the femoral neck trabeculae of patients with OP than those of the control group, while the opposite was observed regarding the levels of LEP. Expression levels of miR-874-3p in hBMSCs were upregulated during osteogenic differentiation, while those of LEP were downregulated. Moreover, miR-874-3p upregulation promoted ALP, RUNX2, OCN, and OSX mRNA expression, cell proliferation, and calcium deposition in hBMSCs. LEP was found to be a target gene of miR-874-3p. Overexpression of LEP inhibited the expression of osteoblast markers and reversed the effect of osteogenic differentiation induced by the upregulation of miR-874-3p. In conclusion, miR-874-3p promoted the proliferation and differentiation of hBMSCs by downregulating the expression of LEP, thus inhibiting OP. miRNAs: microRNAs; OP: osteoporosis; hBMSCs: human Bone Marrow Mesenchymal stem cells; LEP: leptin; DEGs: differentially expressed genes.
微小 RNA（miRNAs）调节成骨分化并影响骨质疏松症（OP）。本研究旨在确定 miR-874-3p 在 OP 中的潜在作用。通过定量逆转录聚合酶链反应（qRT-PCR）测量 35 例 OP 患者和非 OP 患者股骨颈小梁中 miR-874-3p 和瘦素（LEP）的表达水平。通过上调人骨髓间充质干细胞（hBMSCs）中的 miR-874-3p，观察 miR-874-3p 或 LEP 对细胞增殖和碱性磷酸酶（ALP）、 runt 相关转录因子 2（RUNX2）、骨钙素（OCN）和骨形成蛋白 2（OSX）水平的影响。此外，通过茜素红染色（ARS）评估钙沉积水平。还通过生物信息学分析、荧光素酶报告基因测定和 RNA 下拉测定评估 miR-874-3p 和 LEP 影响 hBMSCs 成骨分化的分子机制。OP 患者股骨颈小梁中 miR-874-3p 水平明显低于对照组，而 LEP 水平则相反。hBMSCs 成骨分化过程中 miR-874-3p 表达水平上调，而 LEP 表达水平下调。此外，miR-874-3p 上调促进 hBMSCs 中 ALP、RUNX2、OCN 和 OSX mRNA 表达、细胞增殖和钙沉积。发现 LEP 是 miR-874-3p 的靶基因。LEP 过表达抑制成骨细胞标志物的表达，并逆转 miR-874-3p 上调诱导的成骨分化效应。总之，miR-874-3p 通过下调 LEP 的表达促进 hBMSCs 的增殖和分化，从而抑制 OP。miRNAs：微小 RNA；OP：骨质疏松症；hBMSCs：人骨髓间充质干细胞；LEP：瘦素；DEGs：差异表达基因。
