Osteoporosis is a bone disease characterized by low bone mineral density (BMD) and impaired bone microarchitecture due to the abnormal activity of osteoclasts. Cathelicidins are antimicrobial peptides present in the lysosomes of macrophages and polymorphonuclear leukocytes. LL-37, a cathelicidin, induces various biological effects, including modulation of the immune system, angiogenesis, wound healing, cancer growth, as well as inflammation, and bone loss. A previous study reported direct involvement of LL-37 suppressing osteoclastogenesis in humans. Here, we examined the role of LL-37 in the treatment of osteoporosis using an ovariectomy (OVX) rat model. Our results showed that LL-37 significantly reduced bone loss and pathological injury in OVX rats with osteoporosis. Furthermore, we found that LL-37 significantly increased the activity of the Wnt/β-catenin pathway in OVX rats with osteoporosis, including the increased expression of β-catenin, Osterix (Osx), and Runt-related transcription factor 2 (Runx2), whereas XAV-939, an inhibitor of the Wnt/β-catenin pathway, significantly blocked the effects of LL-37 on bone loss and abnormal bone metabolism. Altogether, our findings suggested that LL-37 exerted a protective role in regulating bone loss and abnormal bone metabolism in rats with osteoporosis by activating the Wnt/β-catenin pathway.