Effects of Gonadotropin-Releasing Hormone Analogues on Ovarian Function and Embryogenesis: A Cyclophosphamide-Induced Mouse Model Study

IF 4.7 1区医学 Q1 OBSTETRICS & GYNECOLOGY

Bjog-An International Journal of Obstetrics and Gynaecology Pub Date : 2025-02-19 DOI:10.1111/1471-0528.18098

Qiwang Lin, Mingzhu Cao, Zijin Xu, He Fei, Yunfeng Jin, Jianqiao Liu, Hua Jiang

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引用次数: 0

Abstract

Objective

To clarify the protective effects of gonadotropin-releasing hormone analogues (GnRHas) on cyclophosphamide (CTX)-induced oocyte number loss and development of potential damage.

Design

Mice model study.

Setting

Laboratory-based animal study conducted in controlled research facilities.

Population

Female C57/BL6 mice subjected to CTX-induced ovarian damage.

Methods

The effects of GnRHa on CTX mice were evaluated in terms of hormones, oocyte count on slices, oocyte count in established three-dimensional–constructed ovaries, in vitro fertilisation, RNA sequencing and microinjection.

Main Outcome Measures

The main outcome measures were the number of oocytes in intact mouse ovaries and oocyte quality, evaluated using three-dimensional (3D) tissue-clearing methods, oxidative stress markers (reactive oxygen species [ROS] and malondialdehyde [MDT]), mitochondrial function (ATP levels), and embryogenesis rates at the two-cell, four-cell and blastocyst stages.

Results

In CTX mice, GnRHa pretreatment did not protect endocrine hormone changes, but protected loss of oocyte number on slice counting. A tissue-clearing technique, CUBIC (Clear, Unobstructed Body Imaging Cocktails), was a suitable method for ovaries clearing, and a 3D method for oocyte counting was validated with accuracy of 105.22% ± 3.48%. By this method, GnRHa was also found to protect the loss of oocyte number (597 ± 28 vs. 222 ± 15, p < 0.0001), which may be mediated by upregulated anti-Müllerian hormone (AMH) levels inhibiting primordial follicle development approved by in vitro culture of ovaries. GnRHa also increased the number of retrieved oocytes in CTX mice (19.4 ± 2.1 vs. 15.0 ± 1.6, p < 0.0001) and developmental ability of oocytes (65.0 ± 4.6 vs. 48.1 ± 4.2 for blastocyst, p < 0.0001). RNA sequencing revealed GnRHa pretreatment downregulated pathways of exogenous drug metabolism, oxidative stress and cytochrome P450, validated by detection of adenosine triphosphate (ATP), MDA and ROS levels. The up-expression of Cox17 (cytochrome c oxidase copper chaperone 17) after GnRHa pretreatment was confirmed by PCR and microinjection of siCox17 increased the embryogenesis from CTX mice.

Conclusions

GnRHa was associated with reduced oocyte loss and improved embryogenesis, likely mediated by AMH and Cox17 upregulation.

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促性腺激素释放激素类似物对卵巢功能和胚胎发生的影响：环磷酰胺诱导的小鼠模型研究》。

目的：阐明促性腺激素释放激素类似物（GnRHas）对环磷酰胺（CTX）诱导的卵母细胞数量减少和潜在损伤发生的保护作用。设计：小鼠模型研究。环境：在受控研究设施中进行的基于实验室的动物研究。群体：ctx诱导卵巢损伤的雌性C57/BL6小鼠。方法：从激素、切片卵母细胞计数、建立的三维构建卵巢卵母细胞计数、体外受精、RNA测序和显微注射等方面评价GnRHa对CTX小鼠的影响。主要结局指标：主要结局指标为完整小鼠卵巢中的卵母细胞数量和卵母细胞质量，使用三维（3D）组织清除方法、氧化应激标志物（活性氧[ROS]和丙二醛[MDT]）、线粒体功能（ATP水平）以及二细胞、四细胞和囊胚期的胚胎发生率进行评估。结果：GnRHa预处理对CTX小鼠内分泌激素变化无保护作用，但对卵母细胞切片计数损失有保护作用。组织清除技术CUBIC （Clear, unstructed Body Imaging Cocktails）是卵巢清除的一种合适方法，验证了卵母细胞计数的3D方法，准确率为105.22%±3.48%。通过该方法，GnRHa还被发现可以保护卵母细胞数量的减少（597±28比222±15），p结论：GnRHa与减少卵母细胞损失和改善胚胎发生有关，可能是由AMH和Cox17上调介导的。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bjog-An International Journal of Obstetrics and Gynaecology 医学-妇产科学

CiteScore

10.90

自引率

5.20%

发文量

345

审稿时长

3-6 weeks

期刊介绍： BJOG is an editorially independent publication owned by the Royal College of Obstetricians and Gynaecologists (RCOG). The Journal publishes original, peer-reviewed work in all areas of obstetrics and gynaecology, including contraception, urogynaecology, fertility, oncology and clinical practice. Its aim is to publish the highest quality medical research in women''s health, worldwide.