Type 2 diabetes mellitus (T2DM) accounts for approximately 90% of all diabetic patients, and osteoporosis is one of the complications during T2DM process. ATP6V1H (V-type proton ATPase subunit H) displays crucial roles in inhibiting bone loss, but its role in osteogenic differentiation remains unknown. Therefore in this study, we aimed to explore the biological role of ATP6V1H in osteogenic differentiation. OM (osteogenic medium) and HG (high glucose and free fatty acids) were used to induce the MC3T3-E1 cells into osteogenic differentiation in a T2DM simulating environment. CCK8 assay was used to detect cell viability. Alizarin Red staining was used to detect the influence of ATP6V1H on osteogenic differentiation. ATP6V1H expression increased in OM-MC3T3-E1 cells, while decreased in OM+HG-MC3T3-E1 cells. ATP6V1H promoted osteogenic differentiation of OM+HG-MC3T3-E1 cells. Overexpression of ATP6V1H inhibited Akt/GSK3β signaling pathway, while knockdown of ATP6V1H promoted Akt/GSK3β signaling pathway. ATP6V1H overexpression promoted osteogenic differentiation of OM+HG-MC3T3-E1 cells. The role of ATP6V1H in osteogenic differentiation in a T2DM simulating environment involved in Akt/GSK3β signaling pathway. These data demonstrated that ATP6V1H could serve as a potential target for osteogenic differentiation in a T2DM simulating environment.

2型糖尿病（T2DM）约占所有糖尿病患者的90％，骨质疏松症是T2DM过程中的并发症之一。ATP6V1H（V型质子ATPase亚基H）在抑制骨质流失中起关键作用，但其在成骨分化中的作用仍然未知。因此，在这项研究中，我们旨在探讨ATP6V1H在成骨分化中的生物学作用。在T2DM模拟环境中，使用OM（成骨培养基）和HG（高葡萄糖和游离脂肪酸）诱导MC3T3-E1细胞成骨分化。使用CCK8测定法检测细胞活力。茜素红染色用于检测ATP6V1H对成骨分化的影响。ATP- 6V1H表达在OM-MC3T3-E1细胞中增加，而在OM + HG-MC3T3-E1细胞中降低。ATP6V1H促进OM + HG-MC3T3-E1细胞的成骨分化。ATP6V1H的过表达抑制了Akt /GSK3β信号通路，而ATP6V1H的敲低则促进了Akt /GSK3β信号通路。ATP6V1H过表达促进OM + HG-MC3T3-E1细胞的成骨分化。在涉及Akt /GSK3β信号通路的T2DM模拟环境中，ATP6V1H在成骨分化中的作用。这些数据表明，ATP6V1H可以作为T2DM模拟环境中成骨分化的潜在靶标。在涉及Akt /GSK3β信号通路的T2DM模拟环境中，ATP6V1H在成骨分化中的作用。这些数据表明，ATP6V1H可以作为T2DM模拟环境中成骨分化的潜在靶标。在涉及Akt /GSK3β信号通路的T2DM模拟环境中，ATP6V1H在成骨分化中的作用。这些数据表明，ATP6V1H可以作为T2DM模拟环境中成骨分化的潜在靶标。