摘要
背景:阿胶强骨口服液治疗骨质疏松疗效肯定,但其确切作用机制有待探讨。目的:观察阿胶强骨口服液含药血清对胎鼠成骨细胞中骨保护素及保护素配体表达的影响,探讨阿胶强骨胶囊治疗骨质疏松的分子水平作用。设计:完全随机分组,对照实验。材料:实验于2003-06/2004-10在华中科技大学同济医学院附属协和医院中西医结合骨代谢实验室完成。选用3月龄Wistar大鼠(雌雄各半)30只,随机分为3组,阿胶强骨口服液组和雌激素组及对照组,每组10只。选用清洁级新生SD大鼠12只用于成骨细胞的分离和培养。方法:①各组Wistar大鼠灌胃7d后,制备各组的含药血清。②将清洁级新生SD大鼠分离的颅骨成骨细胞,制成单细胞悬液,消化传代,取二代培养的成骨细胞,制成细胞悬液。培养细胞被分为5组,分别加同体积药液。阿胶强骨口服液组分别加入用培养液稀释为100和500及1000g/L浓度的阿胶强骨口服液含药血清;雌激素组分别加入100及1000g/L替勃龙含药血清;对照组仅加培养液。同时各组再加入含100g/L小牛血清的培养液,继续培养。③采用四甲基偶氮唑盐比色分析法、3H-胸腺嘧啶核苷掺入法测定成骨细胞增殖,用放免法测定细胞内骨钙素含量,反转录聚合酶链反应测定胎鼠成骨细胞中骨保护素及保护素配体mRNA的表达。④组间比较采用单因素方差分析。主要观察指标:胎鼠成骨细胞经阿胶强骨口服液或利维爱含药血清干预后,细胞中骨保护素及保护素配体mRNA的表达。结果:①成骨细胞增殖四甲基偶氮唑盐比色分析法及3H-胸腺嘧啶核苷测定结果:阿胶强骨口服液组和替勃龙组各药物血清组明显高于对照组(P<0.05～0.01),并在500g/L时达到最大效应(P<0.01),当浓度大于500g/L时,其作用趋于饱和。各浓度阿胶强骨口服液及替勃龙含药血清均可增加成骨细胞骨钙素含量(P均<0.05)。②骨保护素基因mR
BACKGROUND：The therapeutic effects of donkey-hide glue reinforcing bone oral solution on osteoporosis have been determined, but the exact effective mechanism is to be approached. OBJECTIVE： To investigate the effects of donkey-hide glue reinforcing bone oral solution （DGRBOS） medicated serum on osteoprotegerin （OPG）and its ligand（OPGL）mRNAexpression of osteoblast in fetal rats and explore the molecular mechanism of treating osteoporosis with DGRBOS. DESIGN： A randomized controlled trial. MATERIALS： The experiment was carried out from June 2003 to October 2004 in Bone Metabolic Laboratory of Department of Integrative Chinese and Western Medicine, Affiliated Hospital of Tongji Medical College,Huazhong University of Technology and Science. Totally 30 3-month-old Wistar rats （15 males and 15 females） were randomly divided into 3 groups, I.e. DGRBOS group, estrogen group and control group, with 10 rats in every group. 12 clean newborn SD rats were selected to isolate and cul ture osteoblast. METHODS： ①After intragastric administration for 7 days, medicated serum was prepared respectively from the three groups. ②Skull osteoblast isolated from newborn SD rats was made into single cell suspension, then after digestion and passage, the subcultured osteoblast cell was made into cell suspension. The cultured osteoblasts were divided into 5 groups and given equal volumes of drug liquor. The DGRBOS group was given DGR BOS-medicated serum at the concentration of 100, 500 and 1 000 g/L which was diluted by nutrient solution; the estrogen group was given tibolone-medicated serum of 100 and 1 000 g/L; the control group was given only culture fluid. Meanwhile every group was given calf serum （100 g/L） for further culture. ③The osteoblast proliferation was measured by antigenic MTT colorimetric analysis and 3H-TdR penetration method. The in tra-cellular BGP contents were evaluated by radioimmunity .The mRNA expression of OPG and RANKL in osteoblast was analyzed by Rt-PCR. ④ One-way analysis of
出处
《中国临床康复》
CSCD
北大核心
2005年第42期146-148,共3页
Chinese Journal of Clinical Rehabilitation
基金
湖北省卫生厅资助课题(2004-3-130)~~
