摘要
目的:探讨降钙素基因相关肽作用下成骨细胞内蛋白质表达谱的变化。方法:传代培养MG63成骨细胞,随机分为CGRP组和对照组。提取各组成骨细胞内蛋白质,经双向电泳分离、考马斯亮蓝染色后扫描分析蛋白质表达变化,选取6个差异显著的蛋白点进行胶内酶切、肽质量指纹图谱分析和IPI数据库检索。结果:双向电泳可分离(967±17)个蛋白质,点匹配率为(85.1±1.4)%,成骨细胞6种蛋白点表达出现明显差异,与对照组比较,CGRP组6种蛋白质表达下调。质谱分析鉴定出与核糖体定位和合成的新蛋白转位有关的核糖体结合蛋白p180(p180/ribosomereceptor)及影响细胞内钙浓度的钙结合蛋白HRNRHornerin等5种蛋白质。结论:CGRP作用于成骨细胞后,成骨细胞蛋白质组变化显著,涉及与钙离子浓度变化及新蛋白转位有关的几组蛋白质,提示CGRP可能通过这几种蛋白参与第二信使及细胞因子的调节并发挥功能。
Objective：To explore proteomic changes in osteoblasts induced by calcitonin gene-related peptide（CGRP）.Methods：Cultured MG63 osteoblasts in exponential phase of growth were randomly divided into two groups：Cells in CGRP group were treated with CGRP at 10-710-10 mol/L.Those in the control were cultured without CGRP.After protein extraction,2-D electrophoresis（2-DE）was employed to separate proteins,and 6 protein spots expressed differentially were picked out and subjected to tryptic in-gel digest and matrix——assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF）for identification.Results：967±17 proteins were detected and the match rate was（85.1±1.4）%.6 protein points displayed significant changes in CGRP treated osteoblasts,2 decreased in expression and 4 disappeared.5 proteins identified after mass spectrometry including p180/ribosome receptor associated with ribosome fixing and new protein transposition,HRNR Hornerin associated with Ca2＋ accommodation,and so on.Conclusion：The proteome in osteoblasts induced by CGRP alters dramatically,decreased and disappeared protiens are related to the changes of Ca2＋ accommodation,ribosome fixing and new protein transposition in osteoblasts.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2010年第6期705-710,共6页
Journal of Practical Stomatology
基金
国家自然科学基金项目(编号:30973334
30772436)
关键词
降钙素基因相关肽
成骨细胞
蛋白质组
双向电泳
质谱
Calcitonin gene-related peptide； Osteoblasts； Proteome； Two-dimensional electrophoresis； Mass spectrometry
