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卵胞浆内单精子注射女性患者卵丘细胞和卵泡液中与炎症相关的氧化应激和亚硝化应激标志物评估:一项前瞻性研究

Evaluation of Oxidative and Nitrosative Stress Markers Related To Inflammation in The Cumulus Cells and Follicular Fluid of Women Undergoing Intracytoplasmic Sperm Injection: A Prospective Study.

作者信息

Debbarh Hasnae, Jamil Malak, Jelloul Hasnae, Kabit Amal, Ennaji Mohamed, Louanjli Noureddine, Cadi Rachida

机构信息

Department of Biology, Laboratory of Molecular Genetic Physiopathology and Biotechnology, Ain Chock Faculty of Sciences, Hassan II University, Casablanca, Morocco. Email:

Department of Biology, Laboratory of Molecular Genetic Physiopathology and Biotechnology, Ain Chock Faculty of Sciences, Hassan II University, Casablanca, Morocco.

出版信息

Int J Fertil Steril. 2024 Feb 2;18(2):108-114. doi: 10.22074/ijfs.2023.559526.1342.

Abstract

BACKGROUND

Oxidative/nitrosative stress in the oocyte microenvironment could have an impact on intracytoplasmic sperm injection (ICSI) outcomes. The presence of reactive oxygen species (ROS) and reactive nitrogen species (RNS) can stimulate the secretion of pro-inflammatory cytokines, leading to chronic inflammation and potentially affecting embryo as well as oocyte quality. This study aimed to examine the relationship of lipid peroxidation [measured by the malondialdehyde (MDA) assay] with protein carbonyl [measured by the 2,4 dinitrophenylhydrazine (DNPH) assay] levels in cumulus cells (CCs), as well as nitric oxide (NO), peroxynitrite (ONOO), and C-reactive protein (CRP) levels in follicular fluid (FF). The potential relationship of these levels with ICSI outcome was also evaluated.

MATERIALS AND METHODS

In this prospective study, 63 FF samples and their corresponding CCs were collected for ICSI procedures. Spectrophotometry was used to assess levels of DNPH, MDA, NO, and ONOO. CRP levels were evaluated using an immunoturbidimetric assay.

RESULTS

The patients under 37 years with normal ovarian reserve had significantly lower levels of MDA, DNPH, NO, ONOO, and CRP compared to those over 37 years. Additionally, we observed higher levels of MDA, DNPH, NO, ONOO, and CRP in the group with an oocyte maturity rate of less than 60%. No significant difference was observed between the DNPH levels and factors such as infertility duration, embryo quality, pregnancy, or the number of retrieved oocytes. A higher level of MDA, NO, ONOO, and CRP was found to be significantly related to the lower number of retrieved oocytes, longer periods of infertility, poor embryo quality, and negative pregnancy outcomes.

CONCLUSION

Oxidative/nitrosative stress, linking to inflammation in the oocyte microenvironment, can be considered as a potentially useful biomarker for assessing the development and competence of oocytes and embryos and predicting ICSI outcomes.

摘要

背景

卵母细胞微环境中的氧化/亚硝化应激可能会影响卵胞浆内单精子注射(ICSI)的结果。活性氧(ROS)和活性氮(RNS)的存在会刺激促炎细胞因子的分泌,导致慢性炎症,并可能影响胚胎以及卵母细胞的质量。本研究旨在探讨卵丘细胞(CCs)中脂质过氧化水平[通过丙二醛(MDA)测定法测量]与蛋白质羰基水平[通过2,4-二硝基苯肼(DNPH)测定法测量]之间的关系,以及卵泡液(FF)中一氧化氮(NO)、过氧亚硝酸盐(ONOO)和C反应蛋白(CRP)水平之间的关系。还评估了这些水平与ICSI结果之间的潜在关系。

材料与方法

在这项前瞻性研究中,收集了63份用于ICSI程序的FF样本及其相应的CCs。采用分光光度法评估DNPH、MDA、NO和ONOO的水平。使用免疫比浊法评估CRP水平。

结果

与37岁以上的患者相比,卵巢储备功能正常的37岁以下患者的MDA、DNPH、NO、ONOO和CRP水平显著较低。此外,我们观察到卵母细胞成熟率低于60%的组中MDA、DNPH、NO、ONOO和CRP水平较高。DNPH水平与不孕持续时间、胚胎质量、妊娠或回收的卵母细胞数量等因素之间未观察到显著差异。发现较高水平的MDA、NO、ONOO和CRP与回收的卵母细胞数量较少、不孕时间较长、胚胎质量较差和妊娠阴性结果显著相关。

结论

与卵母细胞微环境中的炎症相关的氧化/亚硝化应激可被视为评估卵母细胞和胚胎发育及能力以及预测ICSI结果的潜在有用生物标志物。

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