ABSTRACT Sodium Dehydroacetate (Na‐DHA), a widely used food additive, has raised concerns about the chronic health risks associated with long‐term exposure. However, the potential impact of Na‐DHA on bone metabolism, its contribution to osteoporosis risk, and the specific molecular mechanisms remain unclear. This study aims to systematically elucidate the molecular mechanisms through which Na‐DHA induces osteoporosis by integrating network toxicology, molecular docking, and in vitro experiments. Potential targets of Na‐DHA were identified through multi‐database screening. Osteoporosis‐related genes were extracted from the GEO database (GSE156508) and subjected to differential and enrichment analyses. Common targets between Na‐DHA and osteoporosis were identified using a Venn diagram. A protein–protein interaction (PPI) network was constructed using STRING, and core targets were selected through random forest analysis. Molecular docking of core targets with Na‐DHA was performed using AutoDock Vina. Human bone marrow mesenchymal stem cells (hBMSCs) were used as a model to assess cell viability using the CCK‐8 assay, observe osteogenic/adipogenic differentiation phenotypes through Alizarin Red S and Oil Red O staining, and validate the expression of core targets and osteogenic genes by qRT‐PCR and Western blot. Multi‐database screening identified 325 potential Na‐DHA targets and 500 osteoporosis‐related differential genes. Of these, 34 common key targets were identified, which were mainly enriched in pathways related to lipid metabolism, autophagy, and steroid biosynthesis. Random forest analysis identified LCMT1, ARHGEF11, and VCAM1 as core targets, and molecular docking revealed potential binding interactions between Na‐DHA and all three targets. In vitro experiments demonstrated that 10 μM Na‐DHA significantly inhibited hBMSCs viability, reduced calcium deposition in Alizarin Red S staining, increased lipid droplet accumulation in Oil Red O staining, and downregulated the expression of key osteogenic genes ( BGLAP , SP7 , RUNX2 , ALPL ). Moreover, Na‐DHA significantly upregulated the mRNA and protein expression of LCMT1 and downregulated the mRNA and protein expression of ARHGEF11 and VCAM1. Na‐DHA may increase osteoporosis risk by upregulating LCMT1, downregulating ARHGEF11 and VCAM1, and disrupting lipid metabolism and the balance between osteogenesis and adipogenesis, ultimately disrupting bone metabolic homeostasis. This study provides scientific evidence for assessing the skeletal health risks of Na‐DHA and exploring potential intervention targets for bone toxicity.
Qian et al. (Sun,) studied this question.