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体内及“挽救性体外成熟”人囊胚在有无褪黑素处理情况下的基因表达谱分析

Gene expression profiling of human blastocysts from in vivo and 'rescue IVM' with or without melatonin treatment.

作者信息

Hao Yan, Zhang Zhiguo, Han Dan, Cao Yunxia, Zhou Ping, Wei Zhaolian, Lv Mingrong, Chen Dawei

机构信息

Department of Obstetrics and Gynecology, Reproductive Medicine Center, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230032, P.R. China.

出版信息

Mol Med Rep. 2017 Aug;16(2):1278-1288. doi: 10.3892/mmr.2017.6742. Epub 2017 Jun 9.

Abstract

To evaluate the effect of melatonin supplementation in maturation medium for human 'rescue IVM' and investigate differences in transcriptomic profile of blastocysts developed from oocytes matured in vitro with/without melatonin treatment and in vivo, a total of 314 GV oocytes and 320 MI oocytes were collected from 200 patients younger than 35 years old undergoing ICSI cycle. The oocytes were randomly distributed in the control group (no melatonin) and four other groups of varying melatonin concentrations (10‑11, 10‑9, 10‑7, 10‑5 mol/l). Gene profiling was performed on blastocysts developed from in vivo maturation oocytes (in vivo group), and in vitro maturation (IVM) oocytes with an optimal concentration of melatonin treatment (IVM‑anti group) or without melatonin (IVM group). The ratio of high quality blastocysts was significantly higher in the groups treated with 10‑5 mol/l melatonin compared with others groups. The large‑scale analysis of the transcriptome revealed significant differences in mRNA expression levels. In each group, nine blastocysts were selected for gene expression profiling. The differentially expressed genes were involved in cysteine and methionine metabolism, regulation of apoptotic process, mineral absorption, steroid hormone biosynthesis, Wnt signaling, p53 signaling pathway and other functions. The findings indicated that the IVM procedure may potentially affect DNA methylation and the canonical Wnt signaling pathway. Exogenous melatonin positively influenced quality of blastocysts, which may be mediated via upregulation of p53 signaling and correcting DNA methylation changes caused by 'rescue IVM'. However, this study reflected what was generally referred to as 'rescue IVM' and was not a true reflection of clinical IVM techniques. Therefore, melatonin required further investigation as a promising supplement for use in IVM.

摘要

为评估在人类“挽救性体外成熟(IVM)”的成熟培养基中添加褪黑素的效果,并研究经褪黑素处理与未经处理的体外成熟卵母细胞以及体内成熟卵母细胞发育而来的囊胚转录组谱的差异,我们从200名年龄小于35岁且正在接受卵胞浆内单精子注射(ICSI)周期治疗的患者中收集了总共314个生发泡(GV)期卵母细胞和320个减数分裂中期I(MI)期卵母细胞。这些卵母细胞被随机分配到对照组(不添加褪黑素)和其他四组不同褪黑素浓度(10⁻¹¹、10⁻⁹、10⁻⁷、10⁻⁵mol/L)的组中。对由体内成熟卵母细胞(体内组)、经最佳浓度褪黑素处理的体外成熟(IVM)卵母细胞(IVM-抗组)或未经褪黑素处理的体外成熟卵母细胞(IVM组)发育而来的囊胚进行基因谱分析。与其他组相比,用10⁻⁵mol/L褪黑素处理的组中高质量囊胚的比例显著更高。转录组的大规模分析揭示了mRNA表达水平的显著差异。在每组中,选择9个囊胚进行基因表达谱分析。差异表达基因涉及半胱氨酸和甲硫氨酸代谢、凋亡过程调控、矿物质吸收、类固醇激素生物合成、Wnt信号传导、p53信号通路等功能。研究结果表明,IVM程序可能会潜在影响DNA甲基化和经典Wnt信号通路。外源性褪黑素对囊胚质量有积极影响,这可能是通过上调p53信号传导以及纠正“挽救性IVM”引起的DNA甲基化变化来介导的。然而,本研究反映的是通常所说的“挽救性IVM”,并非临床IVM技术的真实反映。因此,褪黑素作为一种有前景的IVM补充剂,仍需进一步研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d85b/5561793/ef7c89d7def4/MMR-16-02-1278-g00.jpg

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