摘要
目的:建立"种植窗"时期人子宫内膜组织蛋白质差异表达的研究方法,探讨差异蛋白质表达与子宫内膜对胚胎接受性的关系。方法:采用荧光差异凝胶电泳比较"种植窗"前期[黄体生成素(LH)+2 d]和"种植窗"时期(LH+7 d)子宫内膜组织差异表达的蛋白质,并进行蛋白质功能聚类分析并探讨其与胚胎着床关系。Western blotting检测差异蛋白膜联蛋白IV的表达以确认结果。结果:在pH 3~10 NL的双向电泳上显示的胶内差异图像经分析测到(2 555±98)个蛋白质斑点。经鉴定、检索、查找和确认差异表达蛋白质共31个,其中17个上调表达,14个下调表达。查阅文献发现31个差异表达的蛋白质功能涉及到细胞迁移与融合、酶活性改变、信号转导与基因调控、免疫调节、血管生成和凝血纤溶等6个系统,与胚胎着床高度相关。膜联蛋白IV在蛋白质组学和Western blotting表达趋势一致。结论:"种植窗"时期人子宫内膜上皮组织差异表达蛋白质的研究,发现与胚胎着床活动密切相关的6个功能组群蛋白质表达的动态变化,提示它们是参与子宫内膜向接受态转化的重要蛋白质,为探讨"种植窗"时期子宫内膜组织功能变化的研究提供了新的思路和途径。
AIM: To establish a method for determining the differential expression of proteins in human endometrium during the implantation window and to analyze the correlation between altered expression of the proteins 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 the proteomic alternations in the endometria of pre-receptive [day 2 after luteinizing hormone surge(LH+2 d)] state versus the endometria of receptive(LH+7 d) state.Validation of annexin IV was performed by Western blotting. RESULTS: Approximate 2 555±98 polypeptide spots were revealed by densitometry analysis of the 2D protein maps in LH+2 d and LH+7 d endometrial tissues resolved in the linear range of pH 3~10 on the 2D gel,in which 31 proteins were found to be significantly changed,including 17 proteins up-regulated and 14 proteins down-regulated in LH+7 d samples.These 31 identified proteins were classified into 6 functional categories of the correlation with implantation process: cell migration or assimilation,enzymic activity,signal transduction and gene regulation,immunoregulation,vascularization,and blood clotting or fibrinolysis system.The same expression trend of annexin IV was confirmed by Western blotting. CONCLUSION: Human endometrium has a differential proteomic repertoire during the window of implantation.The 6 functional categories of differentially expressed proteins in the receptive endometrium indicate that they play an important role in transforming of the endometrium during the receptive state.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第2期201-206,共6页
Chinese Journal of Pathophysiology
基金
广东省科技计划(No.2009B030801022)
国家自然科学基金资助项目(No.2008-30872762)
关键词
子宫内膜接受性
蛋白质组
促黄体激素
植入
Endometrial receptivity
Proteome
Luteinizing hormone
Implantation