Objective To investigate the effects of glucose and serum deprivation under hypoxia（GSDH）treatment on oxidative stress and apoptosis in rat bone marrow mesenchymal stem cells (BMSCs), so to provide an experimental support for improving the therapeutic efficacy of BMSCs.   Methods The cell injury model was established by hypoxia (1% O2), hypoglycemia (1.0 g/L) and serum deprivation in vitro with extracted and purified rat primary BMSCs. The proliferation ability of BMSCs was detected by colony formation assay, cell cycle assay and CCK-8 assay; the migration ability was observed by wound healing assay and Transwell assay; BMSCs apoptosis was detected by apoptosis assay kit(AnnexinV-FITC/PI), mitochondrial membrane potential assay kit(JC-1) staining and mitochondrial fluorescence probe(Mito-Tracker) staining; Reactive oxygen species (ROS) and calcium ion were used to detect the levels of cellular oxidative stress; Western blotting was conducted to measure the expression level of anti-oxidative stress-related protein glutathione peroxidase 4(GPX4) and the key protein expression of nuclear factor erthroid 2-related factor 2(Nrf2) pathway.   Results Rat primary BMSCs highly expressed CD29 and CD71 and lowly expressed CD45 and CD34; GSDH treatment inhibited cell proliferation (P<0.05) and migration (P<0.05)  of BMSCs, increased ROS and calcium ion levels (P<0.05), and suppressed the protein expression of anti-oxidative stress-related protein GPX4 and GCLC (P<0.01); compared with the blank control group, the apoptosis of BMSCs was significantly increased after GSDH treatment (P<0.05) and the mitochondrial membrane potential (P<0.01) and network size (P<0.01) was reduced. The expression levels of Nrf2 protein as well as the downstream key proteins in the Nrf2 signaling pathway were decreased (P<0.05).   Conclusion The oxidative stress and the further apoptotic damage induced by GSDH treatment in BMSCs are related to the inhibition of Nrf2 signaling pathway.