Objective To investigate the protective effect of lithium chloride (LiCl)-activated Wnt/beta-catenin signaling pathway on glucocorticoid-induced bone loss. Methods C57BL6/J mice were randomly divided into 3 groups,osteoporosis (OP) group and LiCl group were injected intraperitoneally once daily with dexamethasone (50 mg/kg), while control group was injected with normal saline. LiCl group was given LiCl (200 mg/kg) through water administration. After 5 weeks of treatment, mice were sacrificed within 24 h. Anti-active beta-catenin antibody was used in immunohistochemical staining to detect the activity of Wnt/beta-catenin signaling pathway. Bone morphology was observed by HE staining. Meanwhile, bone histomorphometrical parameters were analyzed by microCT. Bone formation and bone resorption of different groups were detected by calcein labeling and TRAP staining, respectively. Results The protein level of active beta-catenin was higher in LiCl group compared with OP group. Bone histomorphometrical parameters including the trabecular bone volume as a percentage of total volume (BV/TV), bone mineral density (BMD), trabecular number (Tb. N) and trabecular space (Tb.Sp) were all significantly improved in LiCl group, even though obvious difference existed between LiCl group and control group (P<0.05). Calcein labeling showed that new bone formation was markedly improved in LiCl group, but was still lower than that in control group. Bone resorption was not significantly changed among three groups as indicated by TRAP staining. Conclusion LiCl administration effectively activates Wnt/β-catenin signaling pathway in vivo, which improve new bone formation and rescue glucocorticoid-induced bone loss.