Department of Orthopedics, Qinghai Provincial People's Hospital, Xining, Qinghai 810007, P.R. China.
Department of Orthopedics, Qinghai Provincial People's Hospital, Xining, Qinghai 810007, P.R. China.
Mol Med Rep. 2022 Sep;26(3). doi: 10.3892/mmr.2022.12808. Epub 2022 Jul 29.
Osteoporosis (OP) is a bone metabolic disease, in which low bone mass and the microarchitectural deterioration of bone tissue contribute to the fragility of bones and increase the risk of fracture. The aim of the present study was to determine the role of microRNA (miR)‑98‑5p in high glucose (HG)‑induced preosteoblasts. HG was used to induce preosteoblasts treated in a differentiation medium to establish an OP model. Next, miR‑98‑5p expression was determined using reverse transcription‑quantitative PCR. Following the transfection of an miR‑98‑5p inhibitor into HG‑treated osteoblasts, cell viability was assessed using a Cell Counting Kit‑8 assay, while alkaline phosphatase (ALP) activity, differentiation ability and the expression of differentiation‑regulated genes osteocalcin and osteopontin were measured using the corresponding ALP, Alizarin red staining, reverse transcription‑quantitative PCR and western blotting assays. The association between miR‑98‑5p and the PI3K/AKT/GSK3β signaling pathway was determined using western blotting. Next, the binding relationship between miR‑98‑5p and bone morphogenetic protein 2 (BMP2) was predicted and verified, and the role of BMP2 in the regulation of the PI3K/AKT/GSK3β signaling pathway was explored using western blotting. The results revealed that miR‑98‑5p expression was upregulated in HG‑induced osteoblasts, and the inhibition of miR‑98‑5p resulted in enhanced cell viability, alkaline phosphatase activity and differentiation in osteoblasts following HG induction. It was also discovered that miR‑98‑5p inhibition activated PI3K/AKT/GSK3β signaling, while knockdown of BMP2, which binds to miR‑98‑5p, enhanced the activation of this signaling pathway and the differentiation ability of osteoblasts. In conclusion, the findings of the present study suggested that the inhibition of miR‑98‑5p expression may activate PI3K/AKT/GSK3β signaling to promote HG‑induced suppression of preosteoblast viability and differentiation by targeting BMP2, which provides a novel insight into future potential molecular markers for OP treatment.
骨质疏松症 (OP) 是一种骨骼代谢疾病，其特征为低骨量和骨组织微结构恶化，导致骨骼脆弱，骨折风险增加。本研究旨在探讨微小 RNA (miR)‑98‑5p 在高糖 (HG) 诱导的前成骨细胞中的作用。采用 HG 诱导分化培养基处理前成骨细胞，建立 OP 模型。采用逆转录定量 PCR 检测 miR‑98‑5p 的表达。用 miR‑98‑5p 抑制剂转染 HG 处理的成骨细胞后，采用细胞计数试剂盒‑8 法检测细胞活力，采用碱性磷酸酶 (ALP) 活性、分化能力以及分化调节基因骨钙素和骨桥蛋白的表达进行测定，分别采用相应的 ALP、茜素红染色、逆转录定量 PCR 和蛋白质印迹法。采用蛋白质印迹法检测 miR‑98‑5p 与 PI3K/AKT/GSK3β 信号通路的相关性。预测并验证 miR‑98‑5p 与骨形态发生蛋白 2 (BMP2) 的结合关系，采用蛋白质印迹法探讨 BMP2 对 PI3K/AKT/GSK3β 信号通路的调节作用。结果表明，HG 诱导的成骨细胞中 miR‑98‑5p 表达上调，抑制 miR‑98‑5p 可增强 HG 诱导的成骨细胞活力和 ALP 活性及分化。此外，miR‑98‑5p 抑制可激活 PI3K/AKT/GSK3β 信号通路，而靶向 miR‑98‑5p 的 BMP2 敲低可增强该信号通路的激活和成骨细胞的分化能力。综上所述，本研究结果表明，抑制 miR‑98‑5p 表达可通过靶向 BMP2 激活 PI3K/AKT/GSK3β 信号通路，促进 HG 诱导的前成骨细胞活力和分化抑制，为 OP 治疗的潜在分子标志物提供了新的见解。
