Department of Reproductive Medicine, The Fourth Hospital of Hebei Medical University, 12 Jiankang Road, Chang 'an District, Shijiazhuang, 050011, China.
Department of Obstetrics, The Fourth Hospital of Shijiazhuang, Shijiazhuang, 050011, China.
J Assist Reprod Genet. 2024 Feb;41(2):363-370. doi: 10.1007/s10815-023-03001-9. Epub 2023 Dec 11.
In vitro fertilization-embryo transfer (IVF-ET) is a widely used treatment for infertility, with oocyte maturation and quality having a significant impact on oocyte fertilization, embryo development, and fetal growth. Mitochondrial transcription factor A (TFAM) is essential for maintaining the mitochondrial oxidative respiratory chain and supplying energy for oocyte development, fertilization, and embryonic development. In this study, we aimed to examine TFAM expression in women undergoing IVF-ET and assess its impact on the IVF outcomes.
We recruited 85 women who underwent IVF-ET treatment for infertility. On the date of egg collection, granulosa cells were extracted from the clear follicular fluid of the first mature egg using ultrasound-guided needle aspiration. The collected granulosa cells served three purposes: (1) detecting TFAM gene expression in granulosa cells via immunocytochemistry, (2) determining TFAM mRNA expression using reverse transcription-PCR (RT-PCR), and (3) measuring TFAM protein expression through western blotting.
Based on the results, we found that TFAM was localized and expressed in the cytoplasm of granulosa cells, whereas no expression was detected in the nucleus. Granulosa cells exhibited a linear correlation between TFAM mRNA and TFAM protein expression. The study participants were divided into three groups using the ternary method based on relative TFAM mRNA expression thresholds of 33% and 76%: the low-expression group (n = 30), the moderate-expression group (n = 27), and the high-expression group (n = 28). When compared to the other two groups, the moderate expression group exhibited a significantly higher egg utilization rate, 2 pronucleus rate, fertilization rate, and clinical pregnancy rate (P < 0.05).
TFAM was detected in the cytoplasm of human ovarian granulosa cells. Women with moderate TFAM expression demonstrate enhanced outcomes in IVF.
体外受精-胚胎移植(IVF-ET)是治疗不孕不育的常用方法,卵母细胞成熟和质量对卵母细胞受精、胚胎发育和胎儿生长有重要影响。线粒体转录因子 A(TFAM)对于维持线粒体氧化呼吸链和为卵母细胞发育、受精和胚胎发育提供能量至关重要。本研究旨在检测接受 IVF-ET 治疗的女性中 TFAM 的表达情况,并评估其对 IVF 结局的影响。
我们招募了 85 名接受 IVF-ET 治疗不孕不育的女性。在取卵日,使用超声引导下的针吸从第一枚成熟卵子的透明卵泡液中提取颗粒细胞。收集的颗粒细胞有三个用途:(1)通过免疫细胞化学检测颗粒细胞中 TFAM 基因的表达;(2)使用逆转录聚合酶链反应(RT-PCR)测定 TFAM mRNA 表达;(3)通过 Western blot 测定 TFAM 蛋白表达。
根据结果,我们发现 TFAM 定位于颗粒细胞的细胞质中并表达,而在细胞核中未检测到表达。颗粒细胞中 TFAM mRNA 和 TFAM 蛋白表达呈线性相关。根据相对 TFAM mRNA 表达阈值的 33%和 76%,研究参与者采用三分法分为三组:低表达组(n=30)、中表达组(n=27)和高表达组(n=28)。与其他两组相比,中表达组的卵利用率、2 原核率、受精率和临床妊娠率显著更高(P<0.05)。
在人卵巢颗粒细胞中检测到 TFAM。TFAM 中度表达的女性在 IVF 中表现出更好的结局。