Objective: To investigate the effects of the dietary bone supplement consisting of whey protein， leucine， calcium citrate and vitamin D3 on bone performance in low-calcium fed rats. Methods: 40 female SD rats were randomly divided into normal group， low-calcium model group， calcium carbonate group， calcium citrate + VD3 and supplement groups. After administration for 90 days， growth performance， femur biomechanical parameters and femur microstructure， calcium metabolism levels， bone formation and bone resorption markers， myocytic factor Irisin levels were detected. The relative mRNA expression of ALP， OC and key genes were detected by quantitative real-time polymenose chain reaction. Results: Compared with the normal group， the rats in low-calcium model group showed no significant changes in body weight and organ index (P>0.05)，but a highly significant decrease in bone mineral density and maximum load (P<0.01). Compared with the low-calcium model group， the values of femoral bone density， maximum load (P<0.05， P<0.01) and the content of the bone formation markers (alkaline phosphatase and osteocalcin) in serum were significantly increased in the other three groups， the bone dietary supplements not only significantly increased femoral length， femoral dry weight， maximum deflection and femoral dry weight (P<0.05， P<0.01)， but also was superior to calcium citrate+VD3 and calcium carbonate in enhancing bone density， restoring bone microstructure and bone formation markers content (P<0.01). In addition， the bone dietary supplements effectively improved gastrocnemius muscle mass， hamstring rate or forelimb tension in low-calcium fed rats (P<0.05， P<0.01) and enhanced muscle biomechanical effects on bone， which also activated Wnt signaling pathway by increasing muscle secretion of Irisin， up-regulated mRNA expression of Runx2， ALP and OC， stimulated osteoblast proliferation， differentiation and enhanced bone formation and mineralization. Conclusion: Bone dietary supplements increased bone formation by activating the Wnt signaling pathway， inhibited bone mineral loss， and restored bone microstructure， increased bone mineral density and bone strength， and improved osteoporosis. This may be related to it improved muscle mass and Irisin secretion， enhanced the biomechanical effect of muscle on bone.