Osteoporosis is a major worldwide public health problem that poses a great economic burden to society. Puerariae Radix, the dried root of Pueraria lobata (Wild.) Ohwi, has been widely used in Asia. This study investigated the effects of Puerariae Radix (PR) on bone loss in ovariectomized (OVX) mice. C3H/HeN mice (10 weeks old) were divided into sham and OVX groups. The OVX mice were treated with vehicle, 17β-estradiol (E2), PR (oral administration, 250 mg/kg/day) or PR (intraperitoneal administration, 50 mg/kg/every other day) for 6 weeks. Grip strength, uterus weight, serum alkaline phosphatase (ALP), estradiol concentration and osteoclast surface levels were measured. Tibiae were analyzed using microcomputed tomography. There were no significant differences in the degree of grip strength, body weight and uterine weight between OVX group and PR-treated group. As compared with the OVX group, the serum estradiol levels were significantly increased in the PR-treated group. PR (i.p.) significantly preserved trabecular bone volume, trabecular bone number, structure model index and bone mineral density of proximal tibiae metaphysic. The administration of PR lowered serum ALP and osteoclast surface levels in OVX mice, suggesting that PR can reduce the bone turnover rate in mice. The results indicate that the supply of PR can prevent OVX-induced bone loss in mice.