胚胎移植前宫腔液外泌体提取及其对妊娠结局的影响分析  

Extraction of exosome from uterine fluid prior embryo transfer and its effect on pregnancy outcome

作　　者：任雨蒙 贾莹莹 石程[1] 付旻[1] 廖礼铭 郑晓峰[2] 韩红敬[1] REN Yu-meng;JIA Ying-ying;SHI Cheng;FU Min;LIAO Li-ming;ZHENG Xiao-feng;HAN Hong-jing(Reproductive Medicine Center,Peking University People’s Hospital,Beijing 100044;School of Life Sciences,Peking University,Beijing 100871)

机构地区：[1]北京大学人民医院生殖医学中心,北京100044 [2]北京大学生命科学院,北京100871

出　　处：《生殖医学杂志》2021年第8期1008-1014,共7页Journal of Reproductive Medicine

基　　金：北京大学人民医院公司及协会资助课题(2019-Q-01)。

摘　　要：目的探讨胚胎移植前宫腔液中是否存在外泌体以及在胚胎移植前抽吸宫腔液是否影响妊娠结局。方法选取2019年4月至2020年1月在北京大学人民医院接受IVF/ICSI并新鲜胚胎移植的79例妇女作为研究对象(研究组),研究组在胚胎移植前抽吸宫腔液然后进行胚胎移植,利用超速离心法提取所收集宫腔液中的外泌体并验证。按照年龄及获卵数1∶2的配比进行匹配,获得同期行IVF/ICSI-ET助孕的158例患者作为对照组,对照组患者不抽吸宫腔液,直接进行胚胎移植。比较两组患者的妊娠结局并探讨移植前抽吸宫腔液是否影响妊娠结局。结果研究组宫腔液中分离并提取出外泌体,经透射电镜下观察到外泌体为有完整包膜的圆形或椭圆形结构,Western Blot方法检测外泌体标志物ALIX、TSG101和CD81均呈阳性表达。比较两组的妊娠结局,研究组的胚胎着床率(36.6%vs.34.4%)、临床妊娠率(55.7%vs.47.5%)、持续妊娠率(46.8%vs.42.4%)略高于对照组,但均无显著性差异(P>0.05)。经多因素Logistic回归分析显示提取宫腔液不影响妊娠结局,仅黄体期长方案为影响持续妊娠率的独立因素,与持续妊娠率呈正相关[OR=2.783,95%CI(1.046,7.408),P=0.040]。结论体外受精-胚胎移植患者胚胎移植前抽吸宫腔液不会对妊娠结局造成负面影响,且宫腔液中可以检测到外泌体;宫腔液抽吸作为一种无创检查,为胚胎着床失败原因的分析提供了一种新的方法。Objective:To investigate whether there is exosome in uterine fluid and whether aspiration of uterine fluid before embryo transfer affects pregnancy outcome.Methods:A total of 79 patients who underwent IVF/ICSI-ET in Peking University People’s Hospital from April 2019 to January 2020 were selected as the research objects(the study group).In the study group,the uterine fluid of the patients was aspirated before embryo transfer.The exosomes of uterine fluid were extracted by ultracentrifugation and verified.Matched according to age and number of collected oocytes,158 patients who underwent IVF/ICSI-ET for the same period were obtained as the control group.They received embryo transfer directly without aspiration of uterine fluid.The pregnancy outcomes were compared between the two groups,and whether aspiration of intrauterine fluid before transplantation is independently related to pregnancy outcome was explored.Results:The exosomes were isolated and extracted from the uterine fluid in the study group.Under the transmission electron microscope,the exosomes were round or oval,with complete membrane structure.Western blot showed that the exosome markers such as ALIX,TSG101 and CD81 were positively expressed.Comparing the pregnancy outcomes between the two groups,the embryo implantation rate(36.6%vs.34.4%),clinical pregnancy rate(55.7%vs.47.5%)and persistent pregnancy rate(46.8%vs.42.4%)in the experimental group were slightly higher than those in the control group,but the differences were not significant(P>0.05).Multivariate logistic regression analysis showed that extraction of uterine fluid did not affect the outcome of pregnancy,and only GnRH agonist long protocol in luteal phase was an independent factor affecting the pregnancy rate,and was positively correlated with the ongoing pregnancy rate[OR=2.783,95%CI(1.046,7.408),P=0.040].Conclusions:Exosomes can be extracted from the uterine fluid of IVF-ET patients before embryo transfer,and aspiration of uterine fluid before embryo transfer has no negative effect on cl

关 键 词：宫腔液 外泌体 子宫内膜容受性 胚胎着床 妊娠结局 

分 类 号：R711.6[医药卫生—妇产科学]