BACKGROUND:: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is still unclear. OBJECTIVE:: To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts. METHODS:: Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly: normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrically administrated with extract of Gukang prescription and normal saline based on ratʼs body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was collected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, all osteoblasts were divided into two groups and treated with collected serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse transcription-PCR in all groups. RESULTS AND CONCLUSION:: mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor activator of nuclear factor kappa B ligand were dramatically lower in the Gukang group compared with the normal control group (P < 0.01). These findings indicate that Chinese medicine Gukang prescription can modulate the expression of core binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.