Most studies evaluated the energy release rate of cortical bone macrostructure under Mode I, Mode II, and mixed Mode I-II loading conditions. However, testing the macrostructural energy release rate requires an initial crack and recording the applied load and the corresponding crack length in real-time, which may introduce measurement errors and differences with the actual fracture scenarios. To further understand how the energy release rate contributed to the cortical bone fracture characteristics, this study predicted the microstructural energy release rate of cortical bone and then investigated its age-related varitions. The microstructural energy release rate of femoral cortical bone in rats from different ages was back-calculated by fitting the experimental and simulated load–displacement curves under compression load. The trends in the microstructural energy release rate were revealed, and the underlying reasons for the age-related changes were investigated by integrating the discussion on the cortical bone mechanical parameters at various levels obtained from the previous experiment. The predicted microstructural energy release rate of femoral cortical bone in the rats from 1, 3, 5, 7, 9, 11, and 15 months of age were in the range of 0.08–0.12, 0.12–0.14, 0.15–0.19, 0.25–0.28, 0.23–0.25, 0.19–0.22, and 0.13–0.16 N/mm, respectively. The statistical analyses showed the significant differences in the microstructural energy release rate at different ages. The results indicated an increasing trend followed by a decrease from 1 to 15 months of age, and the correlations between microstructural energy release rate and age were significant. The age-related variations in the microstructural energy release rate may be linked to the changes in the microarchitecture, and the fracture load is influenced by the micro-level mechanical parameters. Notably, the age-related trends in microarchitecture and energy release rate were similar. These findings were valuable for understanding the mechanism underlying the weakening mechanical properties of cortical bone microstructure with age from an energy perspective.