中药骨康对成骨细胞分泌胰岛素样生长因子1的影响

Effects of Chinese medicine Gukang on the expression of insulin-like growth factor 1 in osteoblasts

背景:中药骨康在临床上有明确的治疗骨质疏松的疗效。目的:假设中药骨康与雌激素同样具有调节成骨细胞的作用,观察其对成骨细胞胰岛素生长因子1表达的影响。设计、时间及地点:随机对照观察实验,体外细胞因子检测,于2005-09/2007-04在广州中医药大学附属骨伤科医院细胞培养实验室完成。材料:新生24h内的SD乳鼠,用于成骨细胞的分离培养。新西兰大白兔10只,用于制备药物血清。随机分为5组:空白对照组,中药骨康9.6g/kg、4.8g/kg、2.4g/kg含药血清组及雌二醇含药血清组,每组2只,雌雄各半。方法:各组大白兔按体表面积的方法给予中药骨康方的提取药物、雌激素和生理盐水灌胃,连续给药1周。最后一次用药后2h行心脏采血,分离血清。取第3代成骨细胞,经0.25%胰酶消化,将细胞悬液以终浓度1×107L-1接种到24孔培养板中,24h细胞贴壁后换为含10%各组血清的培养液,培养4d后收集培养上清液。主要观察指标:用酶联免疫吸附法检测培养上清液中胰岛素样生长因子1的含量。结果:中药骨康9.6g/kg、4.8g/kg组和雌二醇组胰岛素样生长因子1水平高于空白对照组(P<0.05),中药骨康9.6g/kg组和雌二醇组胰岛素样生长因子1水平高于中药骨康2.4g/kg组(P<0.05),中药骨康9.6g/kg组和雌二醇组之间比较差异无显著性(P>0.05),中药骨康2.4g/kg组与空白对照组比较差异无显著性(P>0.05)。结论:骨康用药量达到一定程度时,在促进成骨细胞生成胰岛素生长因子1方面的能力与雌二醇一致。

BACKGROUND： Chinese medicine Gukang has a clear effect of clinical treatment of osteoporosis. OBJECTIVE： To investigate the effects of Chinese medicine Gukang on the expression of insulin-like growth factor 1 （IGF-1） in osteoblasts at the hypothesis of Chinese medicine Gukang possesses the regulating effect on osteoblasts identical with the estrogen. DESIGN, TIME AND SETTING： A randomized control experiment of cytokine detection in vitro was performed at Cellular Culture Laboratory, Affiliated Orthopedic Hospital of Guangzhou Chinese Traditional Medicine University between September 2005 and April 2007. MATERIALS： Ten SD neonatal rats were used for the separation and culture of osteoblasts, and ten New Zealand rabbits were used to prepare the drug serum and then were divided into 5 groups randomly： normal control group, high-, middle-, low-concentration groups and estradiol group. Each group contained two rabbits of either gender. METHODS： Based on the surface area, the rabbits in the high-, middle-, and low-concentration groups were intragastrically administrated with 9.6, 4.8 and 2.4 g/kg serum containing Gukang. Estradiol group was intragastrically administrated with 15% estradiol solution, while the normal control group was given normal saline for l week. Two hours after the last administration, blood samples were taken from the heart of each rabbit. Then serum was collected. Osteoblasts at passage 3 were digested with 0.25% pancreatin, and the suspension at the final concentration of 1×10^7 L^-1 was inoculated into 24-well culture plates. Cellular attachment was observed 24 hours later, and meanwhile 10% serum containing corresponding drugs were added to the medium. The supernatant collection was performed 4 days later. MAIN OUTCOME MEASURES： Enzyme-linked immunosorbent assay techniques were used to detect the expression of IGF-1 in supernatant. RESULTS： Expression of IGF- 1 in osteoblasts in the serum containing the high and middle concentrations of Gukang and estradiol was increased compared with that in the normal control group （P 〈 0.05）. The serum containing low concentration of Gukang was inferior to the high concentration of Gukang and the estradiol （P 〈 0.05）. IGF- 1 expression was of insignificant differences between the high concentration group and the estradiol group, and between the low concentration group and the normal control group （P 〉 0.05）. CONCLUSION： The serum containing certain concentration of Chinese medicine Gukang can increase the secretion of IGF-1 in osteoblasts, and the effect is the same as that of estradiol.