孕激素通过上调内异症患者在位内膜间质细胞的自噬改善子宫内膜容受性的分子机制

Embryo implantation rate being significantly decreased is a common problem in patients with endometriosis who underwent in vitro fertilization embryo transfer (IVF-ET) treatment, the existing treatments are failed to effectively solve this problem. Our previous study found that placing Mirena to uterine cavity that sustainly release of progesterone before transplantation can significantly improve the implantation rate. Maternal fetal interaction in the intrauterine environment is a prerequisite for the establishment of physiological pregnancy and the failure of pregnancy is associated with abnormal maternal fetal immune regulation. We recently found that progesterone promotes the autophagy of endometrial stromal cells (ESC), which induce the differentiation of CD56+CD16-NK (uNK), and the latter promoted embryo implantation. However, the exact molecular mechanism and its role in embryo implantation is not clear. Therefore, taking advantage of in vitro trials for imitating the uterine cavity maternal fetal dialogue microenvironment in endometriosis and in vivo animal experiments, our study intends to investigate the downstream signal pathways which are involved in high level of ESC autophagy on uNK differentiation, and to further explore whether these uNK cells promote the physiological mechanism of embryo implantation. This project will provide a new direction to improve embryo implantation rate in patients with endometriosis and provide scientific basis for new strategies to improve the success rate of IVF-ET.

胚胎种植率显著下降是内异症患者行体外受精-胚胎移植（IVF-ET）治疗时的常见问题，现有治疗方法未能有效解决这一问题。我们前期研究发现移植前宫腔内放置持续释放孕激素的曼月乐环可以显著提高胚胎种植率。宫腔内环境中母-胎交互对话是建立生理性妊娠的先决条件，妊娠失败多与母-胎免疫调节异常对胚胎产生排斥有关。我们近期发现孕激素通过促进内膜间质细胞（ESC）的自噬诱导CD56+CD16-NK(uNK)分化，后者可促进胚胎种植。但确切的分子机制及其在胚胎种植中的作用尚不清楚。故本课题以体外模拟内异症患者宫腔母胎对话微环境和体内动物实验为研究手段，拟研究孕激素信号诱导的高自噬ESC通过何种下游信号诱导宫腔微环境uNK分化，进一步解析这群uNK细胞是否通过调节ESC生物学行为促进胚胎种植的生理机制。本课题将为提高内异症患者的胚胎种植率提供新的研究方向，为寻找改善IVF-ET成功率的新策略提供科学依据。

子宫内膜异位症（简称内异症，Endometriosis，EMS）是妇科常见疾病，其子宫内膜异位于宫腔之外，常因反复周期性出血而引发持续加重的盆腔粘连、疼痛和不孕等症状。内异症在生育期妇女中的发病率约6-10%。约25%-40%的内异症患者需要通过体外受精-胚胎移植(in vitro fertilization - embryo transfer，IVF- ET) 的途径解决生育问题。我们近期发现孕激素通过促进内膜间质细胞（ESC）的自噬诱导CD56+CD16-NK(uNK)分化，后者可促进胚胎种植。但确切的分子机制及其在胚胎种植中的作用尚不清楚。故本课题以体外模拟内异症患者宫腔母胎对话微环境和体内动物实验为研究手段，拟研究孕激素信号诱导的高自噬ESC通过何种下游信号诱导宫腔微环境uNK分化，进一步解析这群uNK细胞是否通过调节ESC生物学行为促进胚胎种植的生理机制。本课题将为提高内异症患者的胚胎种植率提供新的研究方向，为寻找改善IVF-ET成功率的新策略提供科学依据。我们发现了：（1）人参二醇通过降调节ERα和升调节PRα来提高内异症患者ESC的自噬水平；②、Western blotting显示：PPD处理后，自噬相关分子（微管相关蛋白轻链3B（LC3B-II）， Beclin-1 ）表达上调，提示PPD可以提高ESC的自噬。③、PPD处理ESC后，ERα表达下调，而PRα的表达上调。④、P62表达下调。⑤、ERα的拮抗剂盐酸二羟甲基丙烷（MPP dehydrochloride）导致ESC自噬表达上调。这些数据表明PPD可以降调节ERα的表达，升调节PRα的表达，最终提高ESC的自噬水平。(2) 低剂量人参二醇 (15 mg/kg, 100μl)不影响EMS鼠模型中ERα或PRα的表达、增殖或者子宫内膜的自噬。（3）高剂量人参二醇可以逆转雌激素对鼠异位灶自噬的抑制作用。这些结果为我们进一步研究内异症患者自噬水平调节在胚胎种植中的作用以及选择新的治疗策略提供了科学依据。