Naringenin Restores Osteogenic Differentiation in TNF-α-Treated Bone Marrow Mesenchymal Stem Cells by Targeting AKR1B1

Background Chronic inflammatory microenvironments, driven by cytokines like TNF-α, disrupt bone remodeling by suppressing osteogenic differentiation, contributing to inflammatory bone diseases. While TNF-α inhibitors mitigate inflammation, they lack osteoprotective effects. Naringenin, a citrus flavonoid with antioxidative and anti-inflammatory properties, shows potential but its mechanism in counteracting TNF-α-induced osteogenic suppression remains unclear. This study aimed to elucidate how naringenin rescues osteogenesis in TNF-α-treated hBMSCs and identify its molecular targets. Methods hBMSCs were treated with TNF-α and naringenin. Cell viability and osteogenic differentiation were assessed via Live/Dead staining, CCK-8, ALP/ARS staining, and qRT-PCR. Antioxidant activity was evaluated using free radical scavenging assays, alongside SOD/CAT activity measurements. Bioinformatics analyses identified TNF-α-regulated differentially expressed genes. GO, KEGG, and PPI network analyses highlighted key pathways. Molecular docking (CB-Dock2) predicted naringenin’s binding to target proteins. Results Naringenin reversed TNF-α-induced suppression of osteogenic markers and restored mineralization in BMSCs. Naringenin also scavenged free radicals, reduced ROS, enhanced SOD/CAT activity, and attenuated TNF-α-driven inflammatory cytokines. Bioinformatic analyses revealed TNF-α-induced enrichment of NF-κB and cytokine pathways, while GSEA highlighted suppressed ossification. TNF-α upregulated AKR1B1, an enzyme linked to oxidative stress, which naringenin suppressed. Molecular docking confirmed strong binding between naringenin and AKR1B1 (Vina score: -10). Conclusion Naringenin rescues TNF-α-impaired osteogenesis by targeting AKR1B1, restoring redox balance, and inhibiting NF-κB-mediated inflammation. This positions naringenin as a therapeutic candidate for inflammatory bone disorders. The study identifies AKR1B1 as a novel target for mitigating inflammation-driven bone loss, bridging phytochemical research and clinical orthopedics.