甲状旁腺激素间歇性应用对大鼠成骨细胞分泌组的影响  

The secretome profile in rat osteoblasts intermittently exposed to parathyroid hormone

作　　者：蒋兰兰[1] 姜惠[1] 刘翠萍[1] 张克勤[1] 

机构地区：[1]南京医科大学第一附属医院内分泌科,南京210029

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2008年第1期53-58,共6页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：教育部和人事部留学回国人员科研启动基金资助（项目号分别为“教外司留”[2007]24号和苏人通[2006]52号

摘　　要：目的利用蛋白质组学技术研究甲状旁腺激素（PTH1-34）对大鼠成骨细胞分泌组的影响，为进一步研究PtH促成骨作用机制、寻找新的骨代谢调节因子打下基础。方法将大鼠成骨细胞分为ptH刺激组和对照组分别培养，收集第三周期6h末和24h末的无血清培养液，用透析-丙酮沉淀法富集培养液中的分泌蛋白质，然后通过双向电泳寻找差异蛋白，再通过质谱分析和数据库检索鉴定两组样品中差异蛋白的种类和性质。结果6h末和24h末PTH刺激组的分泌组均有明显改变，初步鉴定其中23个差异表达蛋白，发现它们分别属于细胞骨架蛋白、翻译调控相关蛋白、蛋白降解相关蛋白、信号转导相关蛋白、热休克蛋白／分子伴侣、抗氧化酶、胰岛素样生长因子结合蛋白及与糖、脂代谢相关的酶等。结论PTH间歇性应用可使成骨细胞分泌组发生明显改变，这些蛋白可能与PTH促成骨作用相关，本研究为进一步研究PTH促成骨机理、寻找新的骨代谢调节因子提供了实验依据。Objective We applied proteomic methods to investigate the secretome profiles of rat osteoblasts （ROB） with or without intermittent PTH treating, which would contribute to the investigation of the molecular mechanism of PTH effects on osteoblasts. Methods The cultured ROB were divided into PTH-treated group （Itm） and control group （Ctr）. In the third cycle, the cells were cultured in the serum-free medium, which was collected at the end of 6h and 24h. The secreted proteins in the medium were extracted by dialysis-acetone precipitation, and then separated by two-dimensional gel electrophoresis （2-DE）. Subsequently, Some of the selected protein spots were measured by matrix assisted laser desorption ionization time-of-flight mass spectrometry （MADI-TOF-MS）. Finally, the data obtained from peptide mass fingerprinting （PMF） was used for protein database search. Results The reliable 2-DE images were successfully obtained. There were 34 differential protein spots in 6h group, while there were 35 differential protein spots in 24h group. We selected 8 differential protein spots from Itm 6h and 15 differential protein spot from Itm 24h for MADI-TOF-MS analysis. The majority of the modulated proteins belonged to the following functional categories： cytoskeleton, regulators of translation, protein degradation, signaling transport, heat shock protein/chaperones, antioxidase, IGF binding protein and metabolic proteins etc. Conclusion Intermittent PTH treatment can greatly change the secretome of osteoblasts, and these changes may be the basis of the bone-forming effect of PTH.

关 键 词：甲状旁腺激素 成骨细胞 蛋白质组学 分泌组 双向电泳 

分 类 号：R782.2[医药卫生—口腔医学] R318.17[医药卫生—临床医学]