Department of Obstetrics and Gynecology, University of Tennessee Health Science Center, Memphis, TN, USA.
Department of Surgery, University of Texas Rio Grande Valley, Edinburg, TX, USA.
J Assist Reprod Genet. 2018 Oct;35(10):1831-1841. doi: 10.1007/s10815-018-1260-z. Epub 2018 Jul 25.
To determine whether recombinant AMH (rAMH) could prevent post-transplant follicular depletion by acting on the stemness markers Oct-4, Sox2, and NANOG.
This was an experimental study where 12 ovariectomized nude mice were xenotransplanted with vitrified/warmed ovarian cortex obtained from a pre-pubertal girl and Alzet pumps delivering rAMH, or placebo (control), were inserted intra-abdominally. Previously vitrified/warmed ovarian cortex fragments were transplanted after 7 days and then harvested after 14 days from pump placement. We performed real-time RT-PCR analyses, ELISA for AMH, FSH, and estradiol, histologic measurement of ovarian follicles, and immunohistochemistry for Ki67 and TUNEL. The main outcome measures were serum levels and tissue expression of the parameters under investigation and follicle count.
Serum AMH, FSH, and estradiol reflected post-ovariectomy profiles and were mildly influenced by rAMH administration. Ovarian cortex expression of AMH, AMH-R2, VEGF, GDF9, Oct-4, and Sox2 was lower in rAMH mice than in controls, while NANOG was upregulated. There was a non-significant decrease in primordial follicles after vitrification-warming, and xenotransplantation further decreased this number. There were lower cell replication and depressed apoptosis in the rAMH group.
Administration of recombinant AMH in the peri-transplant period did not protect the initial follicular depletion but decreased apoptosis and cellular activation and regulated stem cell markers' tissue expression. These results aid our understanding of the inhibitory effects of AMH on follicular development and show the benefit of administering exogenous AMH at the time of pre-pubertal ovarian cortex transplant to protect the follicles from pre-activation and premature depletion.
确定重组 AMH（rAMH）是否通过作用于干性标志物 Oct-4、Sox2 和 NANOG 来防止移植后卵泡耗竭。
这是一项实验研究，将 12 只去卵巢裸鼠异种移植经玻璃化/解冻的来自青春期前女孩的卵巢皮质，并在腹腔内插入 rAMH 或安慰剂（对照）的 Alzet 泵。先前的玻璃化/解冻卵巢皮质碎片在 7 天后移植，然后在泵放置 14 天后收获。我们进行了实时 RT-PCR 分析、AMH、FSH 和雌二醇的 ELISA、卵巢卵泡的组织学测量以及 Ki67 和 TUNEL 的免疫组织化学分析。主要观察指标为研究参数的血清水平和组织表达以及卵泡计数。
血清 AMH、FSH 和雌二醇反映了卵巢切除术后的特征，并受到 rAMH 给药的轻微影响。rAMH 组的卵巢皮质 AMH、AMH-R2、VEGF、GDF9、Oct-4 和 Sox2 的表达低于对照组，而 NANOG 则上调。玻璃化-解冻后原始卵泡数量略有减少，异种移植进一步减少了这一数量。rAMH 组的细胞复制减少，凋亡抑制。
在移植期间给予重组 AMH 并不能保护初始卵泡耗竭，但可减少凋亡和细胞激活，并调节干细胞标志物的组织表达。这些结果有助于我们理解 AMH 对卵泡发育的抑制作用，并表明在青春期前卵巢皮质移植时给予外源性 AMH 可保护卵泡免受预激活和过早耗竭。
