BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil 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 intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected 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 al 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 dramatical y 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.% 背景：中药骨康在临床上治疗骨质疏松疗效明确，但其具体作用途径尚不清晰。 目的：假设中药骨康通过调节核内结合因子a1水平，控制其下游基因核因子κB受体活化因子配体和骨保护素表达，起到调控成骨细胞生长发育的作用。 方法：新生24 h内的SD乳鼠用于成骨细胞的分离培养。成年SD雌性大鼠用于制备药物血清，随机分为正常血清组和中药骨康组。2组大鼠按体表面积的方法给予中药骨康方的提取药物和生理盐水灌胃，连续给药1周。最后一次用药后2h行心脏采血，分离血清。原代培养并传至第3代经碱性磷酸酶鉴定取得的大鼠成骨细胞，消化计数铺板并分为2组，以上述血清处理72 h，MTT法检测成骨细胞的增殖率，ELISA方法检测碱性磷酸酶分泌量并以相应吸光度值进行纠正，运用RT-PCR检测2组成骨细胞核内结合因子α1及其下游核因子κB受体活化因子配体和骨保护素mRNA表达情况。 结果与结论：中药骨康组成骨细胞骨保护素和核内结合因子a1 mRNA水平显著高于正常血清组，核因子κB受体活化因子配体蛋白和mRNA水平显著低于正常血清组(P <0.01)。实验结果证实，中药骨康可通过影响核内结合因子a1表达，调控其下游基因核因子κB受体活化因子配体和骨保护蛋白表达和分泌，进而发挥治疗骨质疏松的作用。
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