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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 investigate the relationship between endometrial annexin IV protein expression and endometrial receptivity in implantation window. Methods The endometrial pre-implantation window (LH) of women with normal fertility was identified by 2D-DIGE and MALDI-TOF-MS. (LH + 2d) on day 2 and LH + 7d on the day of implantation, respectively. Immunohistochemical streptavidin-peroxidase-linked immunospot (SP) Westernblot technique was used to detect the levels of estrogen receptor (ER) and progesterone (PR) in endometrium. The localization and semiquantitative expression analysis of Annexin IV in endometrial tissues were also performed. Results The electrophoretogram showed that the isoelectric point was about 5.8 and the relative molecular mass was 36000. The expression of protein spots on LH + 7d increased by 2.12 times compared with that on LH + 2d. The peptide fingerprinting (PMF) Annexin IV; Annexin IV protein expression existed in LH + 2d endometrial epithelial cells, annexin IV protein expression in LH + 7d endometrial epithelium and stromal cells, and LH + 7d expression was significantly increased ( P <0 · 05). Conclusion The differential expression of Annexin IV protein in early planting stage and planting window suggests that annexin IV may be an important protein transformed from endometrium to embryo in human.