不明原因反复种植失败患者“种植窗”期血清雌二醇、孕酮增高与肿瘤坏死因子受体作用因子3表达异常的关系

The association between increased serum E_2/P and the altered expression of TRAF3 during the“window of implantation”in women with recurrent implantation failure

目的探讨不明原因反复种植失败（RIF）和正常生育妇女（FC）“种植窗”期血清雌二醇（E2）、孕酮（P）、E2／P水平和子宫内膜组织学时相、雌激素受体（ER）、孕激素受体（PR）和肿瘤坏死因子受体作用因子3（TRAF3）蛋白质的表达及其变化差异与反复种植失败的关系。方法2009年10月至2012年3月于中山大学附属第一医院募集34名女性自愿者（反复种植失败患者15例，生育对照组19名）。在黄体生成激素（LH）＋6～LH＋9t1检测血清E2、P水平，微创法取子宫内膜组织；应用Western blot进行TRAF3的蛋白质水平半定量分析，免疫组织化学技术分析ER、PR和TRAF3定位表达与图象半定量分析。结果“种植窗”期血清E2、P水平，子宫内膜腺上皮细胞ER、PR，基质细胞ER、PR在两组间差异无统计学意义，但RIF患者血清E2／P显著高于对照组（P=0．026）免疫组织化学法分析显示TRAF3主要定位于子宫内膜腔上皮和腺上皮细胞，基质细胞表达很弱；免疫组化染色光密度值分析和Western blot半定量结果均显示不明原因RIF组TRAF3表达显著降低（P=0．02）。结论“种植窗”期子宫内膜组织存在TRAF3的定位与半定量表达；E2／P升高可能影响子宫内膜组织局部分子的表达，

Objective To analyze serum level of Estradiol （ E2 ）, Progesterone （ P）, E2/P, endometrial histological dating, Estrogen receptor （ER） ,Progesterone receptor （ER） , and expression of TNF receptor-associated factor 3 （TRAF3） between women with unexplained recurrent implantation failure and fertile controls during the window of implantation, and discuss their association with repeated embryo implantation failure. Methods A total of 34 women （ 15 of them had RIF, 19 of them were fertile controls） were recruited into this study. Endometrial sample was collected during ＂window of implantation＂ by micro biopsies. Western blot analysis and immunohistochemistry of TRAF3 were performed fur localization and semi-quantitative analysis between two groups. Results There were no differences of serum E2, P, endometrial stromal/glandular ER, PR and normality of histological dating during ＂window of implantation＂ between RIF and controls. The serum E2/P was significantly higher in RIF group compared with controls （ P = 0. 026 ）. Positive staining for the TRAF3 was seen in both epithelial ceils and stromal compartments, with a much more intense staining in epithelial Cells than stromal cells. Semi-quantitative results demonstrate a significant decrease of TRAF3 in women with RIF. Conclusion This study demonstrated the expression and localization of TRAF3 in endometrium of women with RIF during implantation window. The higher ratio of E2/P may have an effect on the local endometrial molecular expression.