摘要
背景：成骨细胞分泌的核核因子κB受体活化素配体的主要作用为保持破骨细胞的分化和活性，抑制破骨细胞的凋亡。目的：观察脂联素对成骨细胞核因子κB受体活化素配体表达的影响，拟进一步探讨其对破骨细胞生成的作用。设计、时间及地点：单一样本观察，于2005-08/2006-03在中南大学湘雅二医院代谢内分泌研究所完成。材料：外科手术中弃之5份取正常成人（35～55岁）髂前上棘松质骨由供者自愿提供；人重组脂联素蛋白购自Calbiochem公司。方法：正常成人髂前上棘松质骨体外培养成骨细胞。分别用0，3，10和30mg／L脂联素干预人成骨细胞48h。成骨细胞和CD14＋外周血单核细胞共培养14d，用30mg／L脂联素干预14d，以25μg／L巨噬细胞集落刺激因子＋50μg/L核因子κB受体活化素配体干预14d作为诱导破骨细胞阳性对照。主要观察指标：分别采用荧光定量聚合酶链反应和酶联接免疫吸附剂测定的方法检测核因子κB受体活化素配体的表达；并用脂联素蛋白干预成骨细胞与外周血单核细胞共同培养系统，观察对破骨细胞生成的作用。结果：①脂联素呈剂量和时间依赖性促进人成骨细胞核因子κB受体活化素配体的表达。②脂联素呈时间和剂量依赖性促进人成骨细胞可溶性破骨细胞异化因子蛋白的分泌。③脂联素干预成骨细胞与CD14＋外周血单核细胞共同培养系统可诱导破骨细胞生成。结论：脂联素可通过诱导成骨细胞核因子κB受体活化素配体表达，诱导破骨细胞生成。
BACKGROUND： The main effect of receptor activator of nuclear factor- κB ligand （RANKL） excreted by osteoblasts is to maintain the differentiation and activity of osteoclasts and to inhibit apoptosis of osteoclasts. OBJECTIVE： To observe the effect of adiponectin on the RANKL expression in osteoblasts and to approach the action of adiponectin on the osteoclastogenesis. DESIGN, TIME AND SETTING： Single sample observation, the experiment was performed at the Institute of Endocrinology and Metabolism, The Second Xiangya Hospital of Central South University between August 2005 and March 2006. MATERIALS： The 5 cancellous bone samples of normal adult （35-55 years old） anterior superior iliac spine were discarded in surgery and offered by donor voluntarily. Reconstituted human adiponectin was purchased from Calbiochem. METHODS： We cultured osteoblasts using cancellous bone samples of normal adult anterior superior iliac spine in vitro. Human osteoblasts were incubated with different doses of adiponectin （0,3,10 and 30 mg/L） for 48 hours. Osteoblasts and CD 14＋peripheral blood mononuclear cells（PBMCs） were co-cultured for 14 days, and intervened by 30 mg/L adiponectin for 14 days, then intervened by 25 μ g/L M-SCF＋ 50 μ g/L RANKL for 14 days. The co-cultured cells were used as positive controls for osteoclasts induction. MAIN OUTCOME MEASURES： We determined the expression of RANKL by FQ-PCR and ELISA, intervened osteoblasts and PBMCs co-culture system by adiponcetin and observed its effect on the osteoclastogenesis. RESULTS： Adiponectin promoted human osteoblast RANKL expression in a dose- and time-dependent manner. Adiponectin induced sPANKL production in a dose- and time-dependent manner in osteoblasts. Adiponectin induced the osteoclast formation in the co-culture systems of osteoblasts and PBMCs. CONCLUSION： These findings show that adiponectin incduces osteoclast formation via stimulating RANKL production in osteoblasts.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第24期4647-4650,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
