Objective: To investigate the effects of puerarin on the pathohistochemistry of the femoral head, bone mineral density (BMD), serum calcium and phosphorus levels in osteoporotic rats, and to analyze its regulatory role in the canonical Wnt signaling pathway. Methods: Thirty-six SPF-grade female Wistar rats were used, and 30 of them successfully established the model of osteoporosis rats were randomly divided into sham operation group, model group and puerarin group. The bone mineral density of the femoral head, the levels of calcium and phosphorus in serum, the gray value of Wnt3a, Wnt5a and β-catenin signaling pathway related proteins and the relative expression levels of mRNA in the femoral head were compared among the three groups. Results: Compared with the sham operation group, In the model group, the trabecular bone number and thickness significantly decreased, while the trabecular separation markedly increased (P<0.01). The relative expression levels of Wnt3a, Wnt5a and β-catenin signaling pathway related proteins and mRNA in Wnt signaling pathway were significantly decreased (P<0.05). Compared with the model group, In the puerarin group, trabecular bone number and thickness significantly increased, while trabecular separation decreased (P<0.01). The protein gray value and mRNA relative expression levels of Wnt3a, Wnt5a and β-catenin in femoral head tissue were significantly increased (P<0.05). Conclusion: Puerarin has a therapeutic effect on osteoporotic rats, and it exerts its effect by regulating Wnt3a, Wnt5a, and β-catenin in the classical Wnt signaling pathway.