Mir-142-5p inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells by targeting Lhx8.
Mir-142-5p inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells by targeting Lhx8.
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DOI：
10.1016/j.heliyon.2023.e19878
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发表时间：
2023-09
期刊：
影响因子：
4
通讯作者：
Lu, Sheng
中科院分区：
文献类型：
--
作者：
Du, Yongjun;Zhong, Hui;Yu, Chen;Lv, Yan;Yao, Yueyi;Peng, Zhi;Lu, Sheng
骨质疏松症（OP）是一种常见的全身性骨代谢疾病，发病率较高，是严重的健康危险因素。成骨分化平衡受骨髓间充质干细胞（BMSCs）调节，在OP的发生和进展中发挥关键作用。尽管 LIM 同源盒 8 (Lhx8) 已被确定影响 BMSC 成骨分化，但其在 OP 中的作用和相关机制仍不清楚。在此，我们旨在阐明Lhx8在BMSCs成骨分化中的作用和机制。培养从野生型和 OP Sprague-Dawley 大鼠中分离的 BMSC，并通过流式细胞术和显微镜进行确认。基于双荧光素酶报告基因测定，用 miR-142-5p 模拟物和 miR-NC（阴性对照）转染 BMSC。进行实时定量逆转录聚合酶链反应和蛋白质印迹分析以确定Lhx8在BMSC成骨分化中的作用。 OP 中 Lhx8 表达显着降低，而 miR-142-5p（可能的 Lhx8 调节因子）的表达显着上调。双荧光素酶报告基因检测表明 miR-142-5p 对 Lhx8 发挥直接靶向调节作用。此外，miR-142-5p模拟物在体外显着抑制BMSCs成骨分化以及Lhx8表达，表明miR-142-5p可能通过Lhx8表达调节参与BMSCs成骨分化，并可能作为OP的潜在诊断靶点。总体而言，这些发现表明 miR-142-5p 通过抑制 Lhx8 来抑制 BMSC 成骨分化。这些可能为进一步研究基于miR-142-5p靶向的OP治疗奠定基础。
Osteoporosis (OP), a common systemic bone metabolism disease with a high incidence rate, is a serious health risk factor. Osteogenic differentiation balance is regulated by bone marrow mesenchymal stem cells (BMSCs) and plays a key role in OP occurrence and progression. Although, LIM homeobox 8 (Lhx8) has been identified to affect BMSCs osteogenic differentiation, its roles in OP and the associated mechanism remains unclear. Here, we aimed to elucidate the role and mechanism of Lhx8 in the osteogenic differentiation of BMSCs. BMSCs isolated from wild type and OP Sprague–Dawley rats were cultured and confirmed via flow cytometry and microscopy. Based on dual-luciferase reporter assay, BMSCs were transfected with miR-142-5p mimics and miR-NC (negative control). Real-time quantitative reverse transcription polymerase chain reaction and Western blot analyses were performed to determine the role of Lhx8 in BMSCs osteogenic differentiation. Lhx8 expression was significantly reduced in OP, whereas that of miR-142-5p, a possible Lhx8 regulator, was significantly upregulated. Dual-luciferase reporter assay demonstrated that miR-142-5p exerted a direct targeted regulatory effect on Lhx8. Moreover, miR-142-5p mimics significantly inhibited BMSCs osteogenic differentiation as well as Lhx8 expression in vitro, indicating that miR-142-5p may be involved in BMSCs osteogenic differentiation via Lhx8 expression regulation and may serve as a potential diagnostic target for OP. Overall, these findings indicated that miR-142-5p inhibits BMSCs osteogenic differentiation by suppressing Lhx8. These may serve as a foundation for further studies on OP treatment based on miR-142-5p targeting.
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