Objective：This study aims to investigate the metabolic changes in bone marrow of female rabbits during the development of osteoporosis（OP）. Methods：The rabbit osteoporotic model was created by ovariectomy（OVX）. The loss of bone mineral density（BMD）was evaluated by dual-energy X-ray absorptiometer（DXA）. Metabolomic profiling of bone marrow was conducted through a multiplatform approach including in vivo proton magnetic resonance spectroscopy（1H-MRS）and in vitro liquid chromatography coupled to mass spectrometry（LC-MS/MS）-based analysis. The analysis was conducted before the surgery（baseline），2-month and 5-month after the surgery. Results：BMD in OVX group decreased gradually over time and was significantly lower than the Sham group at 5-month after OVX（2-month，P < 0.05；5-month，P < 0.01）. 1H-MRS analysis showed lactate and lipid in bone marrow were significantly increased in OVX group. Compared to Sham group，choline，creatine，glutamine and hydroxyproline in bone marrow of OVX qroup decreased dramatically. Similarly，in vitro metabolomic analysis of bone marrow showed dramatic metabolic changes as early as 2-month during the development of osteoporosis. In vivo 1H-MRS analysis showed that，compared to the Sham group，amino acids，acylcarnitines，nucleotides and ceramides were significantly altered. Correlation analysis identified a significant negative correlation between BMD and lactate（r=-0.842，P < 0.01）. Significant positive correlations were found between BMD and choline，creatine，glutamine and hydroxyproline. Conclusion：Some complex and diverse changes in bone marrow metabolism was discovered in this study during the development of OP. In vivo analysis of bone metabolism using 1H-MRS might be a potential non-invasive approach for the evaluation of OP.