人正常子宫内膜蛋白质表达图谱的初步建立  

A pilot study in the establishment of normal human endometrium proteomic profile

作　　者：刘海元[1] 郎景和[1] 冷金花[1] 孙大为[1] 朱兰[1] 刘珠凤[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院妇产科,北京100730

出　　处：《生殖医学杂志》2008年第3期192-195,共4页Journal of Reproductive Medicine

基　　金：国家自然科学基金资助项目(30600669)

摘　　要：目的通过双相电泳方法建立正常子宫内膜组织的蛋白质表达图谱。方法收集正常卵泡期子宫内膜组织5份，采用双相电泳建立子宫内膜组织的蛋白质表达谱；利用凝胶染色技术和图像分析软件进行蛋白质点的识别和检测；随机选择5个蛋白质点，应用基质辅助激光解析离子化飞行时间质谱（MALDI-TOF-MS）和生物信息学方法进行蛋白质的鉴定。结果以固相pH梯度（IPG）胶条pH=4～9进行第一相等电聚焦，以12．5％SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）为第二相，成功建立了正常卵泡期子宫内膜组织蛋白质双相电泳图谱，平均含（523±41）个蛋白点，匹配率81．89％。通过MALDI—TOF-MS对5个蛋白质点进行了鉴定。结论成功建立了人正常子宫内膜蛋白质双相电泳图谱，并应用质谱技术进行了部分蛋白质点的鉴定，为进一步构建人子宫内膜蛋白质表达数据库提供了基础，并为研究子宫内膜生理功能和相关疾病提供了一种新的研究方法。Objectives： To establish the protein profile of human endometrium by two-dimensional electrophoresis. Methods： Five normal endometria in proliferative phase were Collected during trans-abdominal hysterectomy. The whole protein extracts from endometrium were seParated using two-dimensional gel electrophoresis （2 DE）. Protein spots were stained by Comassie blue and detected by an image analysis software. Matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and bioinformatics were used to identify the selected five protein spots. Results： 2 DE maps Of human endometrium were obtained using immobilized pH gradient strips with pH value of 4-9 and 12% polyacrylamide gels. Average protein spots were （523 ± 41） and the matching rate was 81.89 %. Five protein spots were successfully identified by mass spectrometry. Conclusions： 2 DE coupled with MALDI-TOF-MS is a useful tool for the human endometrium proteomic profiling. These data will be useful for establishing human endometrium proteome database and provide a new approach in the study of the physiological functions of endometrium and its related diseases.

关 键 词：子宫内膜 双相电泳 基质辅助激光解吸离子化飞行时间质谱 蛋白质组 

分 类 号：R392.3[医药卫生—免疫学] R737.33[医药卫生—基础医学]