Purpose

Intrafollicular fluid (IFF) melatonin plays a decisive role in maintaining granulosa cells’ DNA integrity and protects them against apoptosis. It reduces oxidative stress and improves the oocyte quality with a higher fertilization rate.

Method

This prospective study investigated the antioxidant property of IFF melatonin and its impact on IVF outcome parameters. We also explored the relative expression of five microRNAs (miR-663b, miR-320a, miR-766-3p, miR-132-3p, miR-16-5p) and levels of cell-free DNA (cfDNA) by real-time PCR in unexplained infertile patients. We collected 425 follicular fluid (FF) samples containing mature oocytes from 295 patients undergoing IVF.

Results

Patients were subgrouped based on IFF melatonin concentration (group A ≤ 30 pg/mL, group B > 70 to ≤ 110 pg/mL, group C > 111 to ≤ 385 pg/mL). Our results showed that patients with ≤ 30 pg/mL IFF melatonin levels have significantly higher oxidative stress markers, cfDNA levels, and lower relative expression of miR-663b, miR-320a, miR-766-3p, miR-132-3p, and miR-16-5p compared to other subgroups (p < 0.001). Similarly, they have a low fertilization rate and a reduced number of high-quality day 3 embryos.

Conclusion

Findings suggest that the therapeutic use of melatonin produces a considerable rise in the number of mature oocytes retrieved, fertilization rate, and good-quality embryo selection. Furthermore, miRNA signature enhances the quality of embryo selection, thus, may allow us to classify them as non-invasive biomarkers to identify good-quality embryos.

中文翻译：

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体外受精患者卵泡周围微环境中褪黑素水平和 microRNA (miRNA) 相对表达谱

 目的

卵泡内液 (IFF) 褪黑激素在维持颗粒细胞 DNA 完整性并保护其免于凋亡方面发挥着决定性作用。它可以减少氧化应激并提高卵母细胞质量并提高受精率。

 方法

这项前瞻性研究调查了 IFF 褪黑激素的抗氧化特性及其对 IVF 结果参数的影响。我们还通过实时探索了五种 microRNA（miR-663b、miR-320a、miR-766-3p、miR-132-3p、miR-16-5p）的相对表达和游离 DNA (cfDNA) 水平不明原因不孕患者的 PCR。我们从 295 名接受 IVF 的患者收集了 425 份含有成熟卵母细胞的卵泡液 (FF) 样本。

 结果

根据 IFF 褪黑激素浓度对患者进行亚组（A 组 ≤ 30 pg/mL，B 组 > 70 至 ≤ 110 pg/mL，C 组 > 111 至 ≤ 385 pg/mL）。我们的结果显示，IFF 褪黑激素水平≤ 30 pg/mL 的患者具有显着较高的氧化应激标志物、cfDNA 水平，以及 miR-663b、miR-320a、miR-766-3p、miR-132-3p 和 miR-766-3p 的相对表达较低。 miR-16-5p 与其他亚组相比 ( p < 0.001)。同样，它们的受精率较低，高质量的第 3 天胚胎数量也减少。

 结论

研究结果表明，褪黑激素的治疗​​用途可显着提高回收的成熟卵母细胞的数量、受精率和优质胚胎的选择。此外，miRNA 特征提高了胚胎选择的质量，因此，我们可以将它们分类为非侵入性生物标志物，以识别优质胚胎。