大鼠髁突软骨细胞发育中差异蛋白的表达变化  

Research on differential protein expression of rat developmental condylar chondrocytes

作　　者：江莉婷[1] 谢银银[2] 魏立[3] 朱雅萍[3] 高益鸣[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院口腔科,上海200025 [2]上海交通大学医学院附属瑞金医院上海血液学研究所,医学基因组学国家重点实验室,上海200025 [3]上海市伤骨科研究所,上海200025

出　　处：《中国口腔颌面外科杂志》2013年第1期15-23,共9页China Journal of Oral and Maxillofacial Surgery

基　　金：上海市科学技术委员会医学引导项目(114119a3500)~~

摘　　要：目的:初步建立大鼠髁突软骨细胞发育过程中蛋白表达差异谱,寻找并鉴定其差异表达蛋白。方法:体外培养出生后1、7、14、28 d共4组SD大鼠髁突软骨细胞,应用甲苯胺蓝染色、Ⅱ型胶原免疫组化染色鉴定软骨细胞。提取各组软骨细胞总蛋白,采用iTRAQ标记定量蛋白,2D nano-HPLC和基质辅助激光解吸/电离串联飞行时间质谱(MALDI-TOF-TOF)技术动态、定量观察出生后大鼠髁突软骨细胞发育过程中差异蛋白的表达及其量变情况,所得数据用MASCOT软件处理,筛选样本之间有意义的差异蛋白,运用GO法进行蛋白分类。结果:共鉴定500种差异蛋白,其中137种具有可信度表达,包括15种高可信度表达,相对分子量在7806.24～608459.2,主要功能为参与各类代谢及发育过程、细胞骨架结构、信号转导,响应各类刺激、免疫应答以及细胞间联系及运输等。结论:本研究获得了目前较为全面的大鼠髁突软骨细胞发育中差异蛋白表达谱,并运用质谱技术成功鉴定了差异表达蛋白,为进一步研究髁突软骨细胞内蛋白功能及在颞下颌关节疾病中的改变与作用奠定基础。PURPOSE： To set up differential protein expression profiling of rat condylar chondrocytes during their development, search and identify their characteristics and functions. METHODS： The 1,7,14 and 28 day-old SD rat condylar ehondrocytes were incubated in vitro, identified by toluidine blue stain and type Ⅱ collagen immunohistochemistry reaction. After extracting from rat condylar ehondrocytes of each group, total proteins were labeled with iTRAQ reagents and resolved using 2D nano-high performance liquid chromatograph （HPLC）,protein spots were then identified by mass spectrometry,using matrix-assisted laser desorption ionization-time-of-flight/time-of-fiight technology （MALDI-TOF/TOF）,to quantitatively analyze the differential protein expression of rat condylar chondrocytes during their development.All data were normalized by MASCOT software to search useful differentially expressed proteins, which were then classified by Gene Ontology. RESULTS： We identified 500 differential proteins,corresponding to 137 reliability proteins, 15 high reliability proteins. Their molecular mass values ranged from 7806.24 to 608459.2. The biological functions of a significant proportion of protein were involved in metabolic and developmental processes , as well as participating in different biological functions including structural constituent of cytoskeleton,signal transduction and molecular signaling, response to stimulus, immune response, cell communication and transport and so on. CONCLUSIONS： This study provided the first differential protein expression profiling of rat developmental condylar chondrocytes,which was identified successfully by mass spectrometry.This would lay a foundation to study the protein expression changes and molecular mechanism of chondrocytes in normal condyle or TMJ diseases in the future.Supported by Medical Leading Project of Science and Technology Commission of Shanghai Municipality（114119a3500）.

关 键 词：ITRAQ 髁突 软骨细胞 蛋白质组学 

分 类 号：R782.6[医药卫生—口腔医学]