电针对COH小鼠子宫内膜HOXA10表达时相介导的种植窗期的调控研究

Endometrial implantation phase restricts the clinical pregnacy rate in vitro fertilization and embryo transfer (IVF-ET ). HOXA10 is important during estrogen and progesterone functioning in the endometrium and accurately regulates temporal and spatial of endometrial implantation phase. We have found electro-acupuncture with HCG injection may improve embryo implantation in the previous experiments. Moreover, we found ovarian hyperstimulation changed the expression of HOXA10, and could be regulated by electro-acupuncture. So we hypothesize that electro-acupuncture improve endometrial implantation phase through regulating the expression phase of HOXA10, so as to improve the pregnancy rate”. We use controlled ovarian hyperstimulation preparation of mice with expression of HOXA10 knocked-up and knocked-down through endometrial gene transfection, observe the expression phase of HOXA10 with real-time fluorescence quantitative PCR, Western blot and so on, watch the change of pinopodes by the scanning electron microscope, to clarify that IVF-ET pregnancy low rate is due to the expression of HOXA10 phrase forward and planting window in advance opening and closing; treat with electro-acupuncture on the day of HCG injection, to explore the mechanism of electro-acupuncture increasing pregnancy rate based on the endometrium implanation window adjusted by HOXA10. The results is expected to give a new idea and theory support to IVF-ET and the treatment of infertility, but also can provide the possible therapeutic target for drugs and physical stimulation on such disease.

子宫内膜种植窗是影响体外受精-胚胎移植妊娠率的主要因素，HOXA10是雌孕激素作用于子宫内膜的重要介导者，对种植窗期进行精密的时空调节，前一国家自然科学基金发现HCG注射日介入电针可改善胚胎着床，预实验表明超排卵改变了HOXA10的表达，电针对其有调节作用，据此提出“电针通过调控HOXA10的表达时相来调节种植窗期以提高妊娠率”的假说。采用超排卵小鼠，运用宫腔内基因转染上调和下调HOXA10的表达，应用FQ-PCR、Western blot等连续观察HOXA10表达，扫描电镜观察胞饮突，阐明体外受精-胚胎移植妊娠率低是因HOXA10表达时相前移，种植窗提前开放和关闭所致；HCG注射日行电针治疗，从HOXA10表达时相介导的种植窗期入手，研究电针提高妊娠率的作用机制。研究成果有望为探索体外受精-胚胎移植和不孕症的治疗提供新的研究思路和理论支撑，也可为该类疾病的药物或物理治疗提供潜在治疗靶点。

IVF-ET技术日趋成熟，但仍摆脱不了高排卵率、低妊娠率的难题，结合电针治疗可提高临床妊娠率。正常子宫内膜仅在一个极短的时期内允许胚胎着床，即“种植窗期”，子宫内膜种植窗是影响IVF-ET 妊娠率的主要因素。HOXA10是一种多效的转录调节因子，在胚胎着床及发育、细胞分化与增殖方面具有重要意义,是雌孕激素作用于子宫内膜的重要介导者，对种植窗期进行精密的时空调节。基于HOXA10从时间和空间上调节子宫内膜种植窗的开放和关闭，我们假定：电针可调控COH小鼠子宫内膜HOXA10的表达时相，改变子宫内膜种植窗期，以提高胚胎移植妊娠率。本项目采用控制性超排卵长方案制备模型小鼠，运用基因转染、分子生物学、形态学等技术，观察超排卵后子宫内膜HOXA10表达时相及种植窗期的变化及电针“关元”、“中极”、“三阴交”穴的影响。结果显示：1.控制性超排卵后，与自然周期鼠囊胚交叉种植，COH假孕鼠受孕率低；2.HCG注射日行电针治疗后，临床妊娠率呈现较明显增长趋势；3.控制性超排卵后，HOXA10、整合素αvβ3、LIF蛋白及mRNA表达峰值前移至受孕第3天；4.电针治疗后，子宫内膜HOXA10、整合素αvβ3、LIF蛋白及mRNA表达时相后移，峰值出现在受孕第4天；5.下调HOXA10的表达，可阻断电针效应，HOXA10、整合素αvβ3、LIF蛋白及mRNA表达峰值出现在受孕第5天；6.控制性超排卵后，子宫内膜胞饮突于受孕第3天表达丰富，电针治疗后恢复至第4天表达。上述证据提示：IVF-ET临床妊娠率低与子宫内膜HOXA10表达时相介导的种植窗期前移有关，电针可调控HOXA10的表达时相，改善胚胎着床。研究成果有望为探索IVF-ET和不孕症的治疗提供新的研究思路和理论支撑，也可为该类疾病的药物或物理治疗提供潜在治疗靶点。