The lack of sufficient mechanical stimulation to the human bone, results in disuse osteoporosis. Low-intensity pulsed ultrasound (LIPUS) promotes fracture healing and the treatment of disuse osteoporosis, but its biomechanical mechanism remains unknown. Simulative research on the mechanical effects of LIPUS on disuse trabecular bone and osteoblasts have been performed. The von Mises stress of disuse trabecular bone and osteoblasts obviously increased under LIPUS irradiation. The average von Mises stress of osteoblasts were two orders of magnitude higher under the irradiation of simulant LIPUS than that without LIPUS irradiation, and the von Mises stress of osteoblasts was positively correlated with the amplitude of sound pressure excitation. The results showed that LIPUS irradiation could obviously improve the mechanical micro-environment of trabecular bone and osteoblasts to alleviate the lack of mechanical stimulation. The results of the research can reveal the biomechanical mechanism of LIPUS in the treatment of disuse osteoporosis to some extent and provide theoretical guidance for clinical treatment of disuse osteoporosis through physical methods.