小鼠胚泡植入不同时期子宫内膜组织细胞蛋白质组研究

Differential Proteome Analysis of Endometrium During Blastocast Implantation in Pregnant Mice

利用固相pH梯度双向凝胶电泳(2D-PAGE)分别分离妊娠第3天(3 d)、第5天(5 d)、第7天(7 d)小鼠子宫内膜的总蛋白质;考马斯亮蓝染色、PDQuest 2DE软件分析获得3个不同孕期的蛋白质表达谱;图像分析以5 d的图谱为参考,3 d、7 d与其匹配率分别为68%、61%.在等电点PI 3-10、分子量14.4 KDa-116.0 KDa范围内分离得3 d、5 d、7 d小鼠子宫内膜蛋白点大约分别为713个、736个、673个.选取差异蛋白质点进行MEDLI-TOF质谱分析,查询数据库,获取有意义的蛋白点,分别为载脂蛋白A-I、膜联蛋白A1、谷光甘肽转移酶、丝氨酸蛋白酶抑制剂、烯醇化酶.研究结果表明,在胚泡植入窗口期(5 d),小鼠子宫内膜合成蛋白质的种类和数量明显增加,以适应胚泡植入.子宫内膜表达的蛋白质在调控子宫基质降解和子宫内膜的局部免疫反应方面中扮演着重要角色.

Total proteins of endometrium from mice pregnant for 3,5 and 7 days were analyzed by IPG 2D- PAGE （two-dimensional gel electrophoresis） and imaging analysis. Twenty differential expression protein spots were identified and analyzed by MALDI-TOF-MS. With the PAGE profile of the 5th day as reference, the number of matched protein spots of the 3rd and 7th day was 68G and 61G, respectively. At an isoelectric point of pH 3-10 and a molecular weight of 14. 4-16. 0 kD,713, 736 and 673 protein spots were separated, respectively, from Coomassie Brilliant Blue stained gels of 3, 5 and 7 days in early pregnant mice. By sequence database searching, five of the twenty differential expression protein spots were identified clearly. They were Apolipoprotein A-I, Annexin A1, utathione transferase Omega-1, Serine protease inhibitor A3M and Alpha-enolase. These results indicated that the kinds and quantity of endometrium proteins increased obviously around 5 days after blastocyst implantation in this study and suggested that the expression of these proteins in early pregnant mice （3--7 days） may be involved in regulating the degradation of uterus matrix and the local immune reaction of endometrium.