成骨分化过程取决于机械信号的关键作用。先前的研究发现，频率为 0.5 Hz 的 2,500 微应变 (με) 机械拉伸应变可促进体外成骨。然而，机械应变在细胞和分子水平上影响成骨的机制尚未完全清楚。本研究旨在探讨机械应变对MC3T3-E1细胞成骨分化的作用机制。通过蛋白质组学分析探讨机械应变对蛋白表达的显着影响。生物信息学鉴定了重要的机械敏感蛋白，并使用实时 PCR 研究了基因的表达。双荧光素酶检测揭示了 miRNA 与其靶基因之间的关系。该基因的过表达和下调表达，探讨其在机械诱导成骨分化和转录组学中的作用，揭示了这一过程的进一步机制。蛋白质组学和生物信息学鉴定了一个重要的机械敏感性低表达蛋白ATP13A3，Atp13a3基因的表达也降低。双荧光素酶测定显示 microRNA-3070-3p (miR-3070-3p) 靶向 Atp13a3 基因。此外，Atp13a3的下调表达促进了成骨分化相关基因和蛋白的表达水平，该过程可能是通过肿瘤坏死因子（TNF）信号通路介导的。Atp13a3响应机械拉伸应变来调节成骨分化，TNF Atp13a3调控的信号通路可能参与了这一过程。这些新颖的见解表明 Atp13a3 可能是一种潜在的成骨和骨形成调节剂。版权所有 © 2024。由 Elsevier B.V. 出版。

The process of osteogenic differentiation hinges upon the pivotal role of mechanical signals. Previous studies found that mechanical tensile strain of 2,500 microstrain (με) at a frequency of 0.5 Hz promoted osteogenesis in vitro. However, the mechanism of the mechanical strain influencing osteogenesis at the cellular and molecular levels are not yet fully understood. This study aimed to explore the mechanism of mechanical strain on osteogenic differentiation of MC3T3-E1 cells.Proteomics analysis was conducted to explore the mechanical strain that significantly impacted the protein expression. Bioinformatics identified important mechanosensitive proteins and the expression of genes was investigated using real-time PCR. The dual-luciferase assay revealed the relationship between the miRNA and its target gene. Overexpression and downexpression of the gene, to explore its role in mechanically induced osteogenic differentiation and transcriptomics, revealed further mechanisms in this process.Proteomics and bioinformatics identified an important mechanosensitive lowexpression protein ATP13A3, and the expression of Atp13a3 gene was also reduced. The dual-luciferase assay revealed that microRNA-3070-3p (miR-3070-3p) targeted the Atp13a3 gene. Furthermore, the downexpression of Atp13a3 promoted the expression levels of osteogenic differentiation-related genes and proteins, and this process was probably mediated by the tumor necrosis factor (TNF) signaling pathway.Atp13a3 responded to mechanical tensile strain to regulate osteogenic differentiation, and the TNF signaling pathway regulated by Atp13a3 was probably involved in this process. These novel insights suggested that Atp13a3 was probably a potential osteogenesis and bone formation regulator.Copyright © 2024. Published by Elsevier B.V.