膜联蛋白IV蛋白差异表达与人类子宫内膜胚胎接受性关系的初步研究  

Study of altered expression of annexin IV and human endometrial receptivity

作　　者：李洁[1] 谭真[1] 黎明涛[2] 刘郁林[2] 刘群[1] 顾熊飞[3] 周建中[1] 庄广伦[1] 

机构地区：[1]中山大学附属第一医院生殖医学中心,广州510080 [2]中山大学附属第一医院基础医学院蛋白质组学实验室,广州510080 [3]中山大学附属第一医院生化教研室,广州510080

出　　处：《中华妇产科杂志》2006年第12期803-805,共3页Chinese Journal of Obstetrics and Gynecology

基　　金：广东省自然科学基金资助项目(2003-31691)

摘　　要：目的探讨种植窗时期人类子宫内膜膜联蛋白IV(annexinIV)蛋白的差异表达与子宫内膜胚胎接受性的关系。方法采用胶内差异双向电泳(2D-DIGE)和基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)技术,鉴定生育能力正常的妇女子宫内膜种植窗前期[黄体生成素(LH)峰第2天(LH+2d)]和种植窗期(LH+7d)内膜差异表达蛋白质;应用免疫组化链酶菌抗生物素蛋白-过氧化物酶连接(SP)法和免疫印迹(westernblot)技术,检测子宫内膜雌激素受体(ER)、孕激素(PR)水平,并对子宫内膜组织进行膜联蛋白IV的定位和半定量表达分析。结果电泳图谱显示等电点约5·8,相对分子质量为36000的蛋白斑点在LH+7d的表达较LH+2d增加2·12倍;经鉴定,此蛋白斑点肽指纹图谱(PMF)为膜联蛋白IV;LH+2d子宫内膜上皮细胞存在膜联蛋白IV蛋白表达,LH+7d子宫内膜上皮和基质细胞中均有膜联蛋白IV蛋白表达,且LH+7d的表达量显著增加(P<0·05)。结论种植窗前期及种植窗期膜联蛋白IV蛋白的差异表达,提示膜连蛋白IV可能是人类子宫内膜向胚胎接受态转化的重要蛋白。Objective To explore the association of altered expression of annexin IV in human endometrium during the implantation window and endometrial receptivity. Methods A comparative proteomic strategy, in a combination of two-dimensional fluorescence difference gel electrophoresis （2D- DIGE） and matrix-assisted laser desorption/ionization time of flight mass spectrometry （ MALDI-TOF-MS）, was adopted to search for proteome alternations of pre-receptive （ clay LH ＋ 2 ） versus receptive （ LH ＋ 7 ） endometria. The location and abundance of the identified differentially expressed protein- annexin IV were analyzed by immunostaining and western blot. Results By comparing protein profiles of LH ＋ 2 and LH ＋ 7 samples, we found a protein up-regulated 2. 12 times in LH ＋7 samples, with a relative molecular weight of 36 000 and an isoelectric point near pH 5.8. It was characterized using mass spectrometry and was identified as armexin IV. Immunohistochemical analysis revealed an altered localization of annexin IV - in the epithelia on day LH ＋ 2, and both in the epithelia and stroma cells on day LH ＋ 7. Protein levels of annexin IV were up-regulated on day LH ＋ 7 compared with that on day LH ＋ 2 by Western blot. Integrated optical density of the object （OPTDI） was 46. 249 ±32. 376 and 249. 507 ±31. 959, respectively（P =0. 004）. Conclusions Our study indicates endometrial samples obtained by microbiopsy are available for proteomics studies. It seems possible that the increased expression of annexin IV during the implantation window plays an important role in the morphological differentiation of the uterus to the receptive state.

关 键 词：钙磷脂结合蛋白IV 子宫内膜 胚胎 蛋白质组学 

分 类 号：R714[医药卫生—妇产科学]