Effect of ulipristal acetate and mifepristone at emergency contraception dose on the embryo-endometrial attachment using an in vitro human trophoblastic spheroid and endometrial cell co-culture model
Effect of ulipristal acetate and mifepristone at emergency contraception dose on the embryo-endometrial attachment using an in vitro human trophoblastic spheroid and endometrial cell co-culture model
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DOI：
10.1093/humrep/dex328
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发表时间：
2017-12-01
影响因子：
6.1
通讯作者：
Lee, Kai-Fai
中科院分区：
文献类型：
--
作者：
Li, Hang-Wun Raymond;Li, Ying-Xing;Lee, Kai-Fai
研究问题：醋酸乌利司他（UPA）和米非司酮在相当于紧急避孕（EC）剂量的浓度下是否都能抑制胚胎-子宫内膜附着？总结回答：UPA和米非司酮在浓度对应于EC剂量不具有对胚胎着床的抑制作用，虽然米非司酮在较高浓度似乎有这样的effect.What是已知的：左炔诺孕酮是常用的EC，但它只通过抑制排卵的作用。与左炔诺孕酮相比，UPA和米非司酮作为EC具有更高的功效;虽然有一些提示米非司酮可能干扰着床，但UPA是否具有排卵后抑制着床的作用尚待证实。利用JAr细胞系制成的滋养层球体作为胚胎替代物的体外实验研究，将石川细胞系和原代人子宫内膜细胞培养成单层作为子宫内膜替代物。原代子宫内膜细胞收集从9名志愿者妇女在中期黄体期consent.PARTICIPANTS/MATERIALS，设置，方法：这项研究是在一所大学的妇科单位进行。将JAr和石川细胞系（或原代子宫内膜细胞）用分级浓度的UPA（0、0.04、0.4和4 μ M）或米非司酮（0、0.1、1和10 μ M）处理24小时。使用体外JAr球体-子宫内膜共培养模型研究胚胎-子宫内膜附着。孕激素受体、β-连环蛋白和糖原合成酶激酶3 β的表达（GSK-3 β）分别用实时RT-PCR和Western blotting进行研究。主要结果和机会的作用：在石川实验中，在0 ℃下用UPA处理后，JAr球体附着率没有显著差异（93.0%）、0.04（93.6%）、0.4（93.4%）和4（91.4%）μ M浓度处理后，贴壁率仅在10 μ M浓度下降低（79.8%，P < 0.0001），而在0.1（92.1%）或1.0（95.2%）μ M浓度下不降低。在原代子宫内膜细胞实验中，用UPA 4 μ M处理后的JAr球体附着率（42.6%）与对照组（46.5%）相比也没有显著差异（P > 0.05）。UPA和米非司酮均能显著上调孕激素受体的表达。用UPA 4 μ M或米非司酮10 μ M处理后，β-连环蛋白和GSK-3 β的表达没有显着变化（P > 0.05）.局限性，预防原因：共培养模型只是一种替代物，可能不能完全代表体内胚胎植入的复杂过程，尽管目前还没有完全类似于后者的用于研究体外植入的完美模型。UPA和可能的米非司酮在相应于EC剂量的浓度下对胚胎着床缺乏抑制作用是避孕咨询的重要信息。
STUDY QUESTION: Do both ulipristal acetate (UPA) and mifepristone inhibit embryo-endometrial attachment at concentrations corresponding to the emergency contraception (EC) dose?SUMMARY ANSWER: Both UPA and mifepristone at concentrations corresponding to the EC dose do not have an inhibitory effect on embryo implantation, although mifepristone at a higher concentration appeared to have such an effect.WHAT IS KNOWN ALREADY: Levonorgestrel is commonly used for EC, but it only acts through inhibition of ovulation. UPA and mifepristone have higher efficacy as EC compared to levonorgestrel; while there is some suggestion that mifepristone may interfere with implantation, whether UPA has post-ovulatory action in inhibiting implantation is yet to be confirmed.STUDY DESIGN, SIZE, DURATION: An in vitro experimental study using trophoblastic spheroids made from JAr cell line as the embryo surrogate, and the Ishikawa cell line and primary human endometrial cells cultured to monolayer as the endometrial surrogate. The primary endometrial cells were collected from nine volunteer women in the mid-luteal phase with consent.PARTICIPANTS/MATERIALS, SETTING, METHODS: The study was conducted in a university gynaecology unit. The JAr and Ishikawa cell lines (or primary endometrial cells) were treated with graded concentrations of UPA (0, 0.04, 0.4 and 4 mu M) or mifepristone (0, 0.1, 1 and 10 mu M) for 24 h. Embryo-endometrial attachment was studied using an in vitro JAr spheroid-endometrial co-culture model. Expressions of progesterone receptor, beta-catenin and glycogen synthase kinase 3 beta (GSK-3 beta) were studied with real-time RT-PCR and Western blotting, respectively.MAIN RESULTS AND THE ROLE OF CHANCE: In the Ishikawa experiments, there was no significant difference in the JAr spheroid attachment rate after treatment with UPA at 0 (93.0%), 0.04 (93.6%), 0.4 (93.4%) and 4 (91.4%) ae M concentrations (P > 0.05); the attachment rate was reduced after treatment with mifepristone only at 10 mu M (79.8%, P < 0.0001) but not at 0.1 (92.1%) or 1.0 (95.2%) mu M concentrations. In the primary endometrial cell experiments, again no significant difference was observed in the JAr spheroid attachment rate after treatment with UPA 4 mu M (42.6%) compared to control (46.5%, P > 0.05). Both UPA and mifepristone could significantly up-regulate progesterone receptor expression. There was no significant alteration in expression of beta-catenin and GSK-3 beta after treatment with UPA 4 mu M or mifepristone 10 mu M (P > 0.05).LIMITATIONS, REASONS FOR CAUTION: The co-culture model is only a surrogate which may not fully represent the complicated process of embryo implantation in vivo, although there is no existing perfect model for studying implantation in vitro which fully resembles the latter.WIDER IMPLICATIONS OF THE FINDINGS: The lack of inhibitory effect on embryo implantation by UPA and possibly mifepristone at concentrations corresponding to the EC dose is an important information for contraceptive counseling.