Gametogenesis Research Center, Institute for Basic Sciences, Kashan University of Medical Sciences, Kashan, Iran.
Biotechnology Department, Faculty of Chemistry, University of Kashan, Kashan. Iran.
Reprod Toxicol. 2023 Dec;122:108490. doi: 10.1016/j.reprotox.2023.108490. Epub 2023 Oct 19.
Improving oocyte competence during chemotherapy is widely known as a contributing factor to increasing the probability of fertility. Additionally, the role of cumulus cells in oocyte quality is of utmost importance. Therefore, this study was designed to simultaneously probe into the relative gene expression of oocytes and cumulus cells as biomarkers of oocyte quality with cyclophosphamide and L-carnitine treatment. A total of 60 adult NMRI mice were divided into four groups: control, L-carnitine (LC), cyclophosphamide (CP), and cyclophosphamide+L-carnitine (CP+LC). The relative mRNA expression levels of oocyte quality genes including growth differentiation factor 9 (Gdf9), hyaluronan synthase 2 (Has2), and mitochondrial sirtuin 3 (Sirt3) in oocytes, and genes involved in bilateral communication between cumulus cells and between the oocyte and its neighboring cumulus cells including connexin 37 (Cx37) and connexin 43 (Cx43) were detected by Real-time-PCR. DCFH-DA staining analyzed the level of intracellular ROS in oocytes. Under the influence of L-carnitine, Gdf9, Has2, Cx43, and Cx37 were significantly up-regulated (p ≤ 0.05). However, cyclophosphamide considerably reduced the expression of all these genes (p ≤ 0.05). The expression of the Sirt3 gene in the CP group increased significantly compared to the other groups (p ≤ 0.05). Analysis of fluorescent images revealed that the level of intracellular ROS in the cyclophosphamide group was significantly increased compared to the other groups (p ≤ 0.05), while it plummeted in the L-carnitine group (p ≤ 0.05). L-carnitine as an antioxidant can reduce the destructive effects of cyclophosphamide and enhance bilateral communications between oocytes and cumulus cells, and it may ultimately lead to an increase in the fertility rate.
在化疗过程中提高卵母细胞的能力被广泛认为是提高生育概率的一个因素。此外，卵丘细胞在卵母细胞质量中的作用至关重要。因此，本研究旨在同时探讨卵母细胞和卵丘细胞的相对基因表达作为环磷酰胺和左旋肉碱治疗的卵母细胞质量的生物标志物。将 60 只成年 NMRI 小鼠分为 4 组：对照组、左旋肉碱（LC）组、环磷酰胺（CP）组和环磷酰胺+左旋肉碱（CP+LC）组。通过实时 PCR 检测卵母细胞中包括生长分化因子 9（Gdf9）、透明质酸合酶 2（Has2）和线粒体沉默调节蛋白 3（Sirt3）在内的卵母细胞质量基因以及卵丘细胞之间和卵母细胞与其相邻卵丘细胞之间的双向通讯相关基因的相对 mRNA 表达水平，包括连接蛋白 37（Cx37）和连接蛋白 43（Cx43）。DCFH-DA 染色分析卵母细胞内 ROS 水平。在左旋肉碱的影响下，Gdf9、Has2、Cx43 和 Cx37 的表达显著上调（p≤0.05）。然而，环磷酰胺显著降低了所有这些基因的表达（p≤0.05）。CP 组 Sirt3 基因的表达与其他组相比显著增加（p≤0.05）。荧光图像分析显示，与其他组相比，环磷酰胺组的细胞内 ROS 水平显著升高（p≤0.05），而左旋肉碱组则显著降低（p≤0.05）。左旋肉碱作为一种抗氧化剂可以减少环磷酰胺的破坏作用，增强卵母细胞和卵丘细胞之间的双向通讯，最终可能导致生育能力的提高。
