METHODS: Peripheral blood was collected on the day of ovulation and the 5th day after ovulation for high-throughput sequencing from women who achieved clinical pregnancy through natural cycle frozen embryo transfer. RT-qPCR assessed miR-135a-5p expression in the endometrium tissue or cells during the mouse implantation window or decidualization. Scanning electron microscopy was utilized to observe pinopode morphology and quantity in mice overexpressing miR-135a-5p during the WOI. Human endometrial stromal cells (HESC) and artificial induction of mouse uterine decidualization were used to explore whether miR-135a-5p overexpression inhibits decidualization by regulating HOXA10 and BMPR2. Furthermore, the impact of miR-135a-5p on HESC proliferation and HTR8/SVneo invasion was explored.
RESULTS: A total of 54 women were enrolled in the study. bioinformatics analysis and animal models demonstrated that miR-135a-5p was significantly downregulated during the WOI, and its high expression can lead to abnormal pregnancy outcomes. Overexpression of miR-135a-5p resulted in the absence of pinopode in mouse endometrial tissue during the WOI. High miR-135a-5p levels were found to potentially inhibit endometrial tissue decidualization by downregulating HOXA10 and BMPR2 expression. Finally, CEBPD was identified as a potential regulator of miR-135a-5p, which would explain the decreased miR-135a-5p expression during the WOI.
CONCLUSIONS: MiR-135a-5p expression is significantly downregulated during the WOI. High miR-135a-5p levels suppress pinopode development and endometrial tissue decidualization through HOXA10 and BMPR2, contributing to inadequate endometrial receptivity.
方法:在排卵当天和排卵后第5天采集通过自然周期冷冻胚胎移植获得临床妊娠的女性外周血进行高通量测序。RT-qPCR评估小鼠植入窗口或蜕膜化期间子宫内膜组织或细胞中的miR-135a-5p表达。扫描电子显微镜用于观察WOI期间过表达miR-135a-5p的小鼠中的pinopode形态和数量。人子宫内膜基质细胞(HESC)和小鼠子宫蜕膜化的人工诱导用于探索miR-135a-5p过表达是否通过调节HOXA10和BMPR2抑制蜕膜化。此外,探讨miR-135a-5p对HESC增殖和HTR8/SVneo侵袭的影响。
结果:共有54名女性参加了这项研究。生物信息学分析和动物模型证明miR-135a-5p在WOI过程中显著下调,高表达可导致妊娠结局异常。miR-135a-5p的过表达导致在WOI期间小鼠子宫内膜组织中不存在pinopode。发现高miR-135a-5p水平可能通过下调HOXA10和BMPR2表达来抑制子宫内膜组织蜕膜化。最后,CEBPD被鉴定为miR-135a-5p的潜在调节因子,这可以解释在WOI期间miR-135a-5p表达降低。
结论:MiR-135a-5p表达在WOI期间显著下调。高miR-135a-5p水平通过HOXA10和BMPR2抑制pinopode发育和子宫内膜组织蜕膜化,导致子宫内膜容受性不足。