Du Yongjun, Zhong Hui, Yu Chen, Lv Yan, Yao Yueyi, Peng Zhi, Lu Sheng
Orthopaedics Department, The First People's Hospital of Yunnan Province, The Affiliated Hospital of Kunming University of Science and Technology, The Key Laboratory of Digital Orthopaedics of Yunnan Provincial, Yunnan Provincial Center for Clinical Medicine in Spinal and Spinal Cord Disorders, Kunming, 650034, China.
Medical School, Kunming University of Science and Technology, Kunming, Yunnan 650500, China.
Heliyon. 2023 Sep 9;9(9):e19878. doi: 10.1016/j.heliyon.2023.e19878. eCollection 2023 Sep.
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.
骨质疏松症(OP)是一种常见的、发病率高的全身性骨代谢疾病,是一个严重的健康风险因素。成骨分化平衡由骨髓间充质干细胞(BMSCs)调节,在OP的发生和发展中起关键作用。尽管已确定LIM同源盒8(Lhx8)会影响BMSCs的成骨分化,但其在OP中的作用及相关机制仍不清楚。在此,我们旨在阐明Lhx8在BMSCs成骨分化中的作用和机制。从野生型和OP斯普拉格-道利大鼠中分离出BMSCs并进行培养,通过流式细胞术和显微镜检查进行确认。基于双荧光素酶报告基因检测,用miR-142-5p模拟物和miR-NC(阴性对照)转染BMSCs。进行实时定量逆转录聚合酶链反应和蛋白质免疫印迹分析,以确定Lhx8在BMSCs成骨分化中的作用。Lhx8在OP中的表达显著降低,而作为可能的Lhx8调节因子的miR-142-5p的表达则显著上调。双荧光素酶报告基因检测表明,miR-142-5p对Lhx8发挥直接的靶向调节作用。此外,miR-142-5p模拟物在体外显著抑制BMSCs的成骨分化以及Lhx8的表达,表明miR-142-5p可能通过调节Lhx8的表达参与BMSCs的成骨分化,并且可能作为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BMSCs的成骨分化。这些可能为基于miR-142-5p靶向的OP治疗的进一步研究奠定基础。