Department of Anatomy, Faculty of Medicine, Social Determinants of Health Research Center, Gonabad University of Medical Science, Gonabad, Iran.
Department of Physiology, Faculty of Medicine, Infectious Diseases Research Center, Gonabad University of Medical Sciences, Gonabad, Iran.
BMC Womens Health. 2023 Oct 4;23(1):525. doi: 10.1186/s12905-023-02673-w.
In recent decades, in vitro fertilization (IVF) has been widely used as a method of assisted reproductive technology (ART) to improve fertility in individuals. To be more successful in this laboratory method, we used the presence of two common types of antioxidants (melatonin and vitamin C) simultaneously and exclusively in IVF medium.
The cumulus-oocyte complexes (COCs) were obtained from Gonadotropin-releasing hormone (GnRH) and Human Chorionic Gonadotropin (HMG) -stimulated mice. Subsequently, metaphase II (MII) oocytes were fertilized in vitro. In the experiment, the IVF medium was randomly divided into two equal groups: The control group did not receive any antioxidants. In the treatment group, 100 µM melatonin and 5 mM vitamin C were added to the IVF medium. Finally, oocytes and putative embryos transferred into developmental medium and cultured 120 h after IVF to the blastocyst stage. After and before IVF, oocytes and putative embryos were stained with dichlorodihydrofluorescein diacetate (DCFDA) and the H2O2 level was measured with an inverted fluorescence microscope using ImageJ software. At the end of the fifth day after IVF, the expression of Bax and B cell lymphoma 2 (Bcl2) was evaluated using real-time PCR.
The levels of reactive oxygen species (ROS) in oocytes and putative embryos observed in the treatment group demonstrated a significant reduce compared to the control group (p ≤ 0.01. (.Furthermore, the number of embryos in the blastocycte stage(P < 0.05), the expression level of the Bcl2 (P < 0.05) gene, the Bax unlike gene, significantly increased compared with the control group.
We conclude that the presence of melatonin and vitamin C antioxidants simultaneously and exclusively in the IVF medium leads to a reduction in ROS and ,as a result, improves the growth of the embryo up to the blastocyst stage.
近几十年来，体外受精（IVF）已被广泛用作辅助生殖技术（ART）的一种方法，以提高个体的生育能力。为了使这种实验室方法更加成功，我们同时且专门在 IVF 培养基中使用了两种常见类型的抗氧化剂（褪黑素和维生素 C）。
从促性腺激素释放激素（GnRH）和人绒毛膜促性腺激素（HMG）刺激的小鼠中获得卵丘-卵母细胞复合物（COCs）。随后，体外受精使卵母细胞进行减数分裂 II 期（MII）。在实验中，将 IVF 培养基随机分为两组：对照组不接受任何抗氧化剂。在治疗组中，向 IVF 培养基中添加 100µM 褪黑素和 5mM 维生素 C。最后，将卵母细胞和假定胚胎转移到发育培养基中，并在 IVF 后 120 小时培养至囊胚阶段。在 IVF 前后，使用二氯二氢荧光素二乙酸酯（DCFDA）对卵母细胞和假定胚胎进行染色，并使用倒置荧光显微镜和 ImageJ 软件测量 H2O2 水平。在 IVF 后的第五天结束时，使用实时 PCR 评估 Bax 和 B 细胞淋巴瘤 2（Bcl2）的表达。
与对照组相比，治疗组卵母细胞和假定胚胎中观察到的活性氧（ROS）水平显著降低（p≤0.01）。此外，与对照组相比，囊胚期胚胎数量（P<0.05）、Bcl2 基因表达水平（P<0.05）、Bax 基因表达水平显著增加。
我们得出结论，同时且专门在 IVF 培养基中存在褪黑素和维生素 C 抗氧化剂可降低 ROS 水平，并由此提高胚胎生长至囊胚阶段的水平。
