Du Tianqi, Su Han, Cao Dan, Meng Qingxia, Zhang Ming, Liu Zhenxing, Li Hong
Center for Human Reproduction and Genetics, Affiliated Suzhou Hospital of Nanjing Medical University, Suzhou Municipal Hospital, Gusu School, Nanjing Medical University, Suzhouing, China.
Key Laboratory of Reproductive Medicine and Offspring Health, Affiliated Suzhou Hospital of Nanjing Medical University, Nanjing Medical University, Suzhouing, China.
Eur J Med Res. 2024 Dec 18;29(1):593. doi: 10.1186/s40001-024-02181-z.
Ovarian tissue cryopreservation has become a promising alternative for fertility preservation in cancer patients, allowing ovarian tissue to be stored for future autotransplantation. Oxidative stress damage occurring during the cryopreservation process may impact tissue quality and function. This study aims to investigate the protective effects and potential mechanisms of Mitoquinone (MitoQ), a mitochondria-targeted derivative of the antioxidant ubiquinone, during the vitrification of ovarian tissue in mice.
KGN cells were treated with various concentrations (0.1, 1, 10, and 50 μM) of MitoQ to determine the optimal concentration. Female ICR mice were divided into three groups: control, conventional vitrification, and MitoQ-supplemented vitrification. Ovarian samples were cryopreserved, thawed, and assessed for tissue morphology using Hematoxylin and Eosin (H&E) staining, and mitochondrial changes using immunofluorescence, transmission electron microscopy, and Western blot analysis. RNA sequencing (RNA-seq) was employed to explore potential protective mechanisms. Autotransplantation experiments were conducted, and the long-term effects of MitoQ on ovarian function were evaluated by counting follicle numbers through H&E staining and measuring serum estradiol and AMH levels using ELISA.
MitoQ at 1 μM was found to be the optimal concentration for maintaining follicular morphology after vitrification. It effectively reduced mitochondrial oxidative damage, preserved mitochondrial morphology, and regulated the expression of mitochondrial dynamics proteins (Drp1 and Mfn2). RNA-seq and Western blot analyses revealed that MitoQ inhibited the p38 MAPK pathway, thereby reducing apoptosis. Additionally, autotransplantation experiments showed that MitoQ treatment significantly increased follicle counts, estradiol (E2), and anti-Müllerian hormone (AMH) levels compared to conventional vitrification.
MitoQ effectively mitigates vitrification-induced oxidative damage, maintains mitochondrial homeostasis, and preserves both follicular reserve and endocrine function. These findings suggest that MitoQ is a valuable adjunct in ovarian tissue cryopreservation and could significantly improve fertility preservation outcomes for cancer patients.
卵巢组织冷冻保存已成为癌症患者生育力保存的一种有前景的替代方法，可将卵巢组织储存以备将来自体移植。冷冻保存过程中发生的氧化应激损伤可能会影响组织质量和功能。本研究旨在探讨抗氧化剂泛醌的线粒体靶向衍生物米托醌（MitoQ）在小鼠卵巢组织玻璃化冷冻过程中的保护作用及潜在机制。
用不同浓度（0.1、1、10和50μM）的MitoQ处理KGN细胞以确定最佳浓度。将雌性ICR小鼠分为三组：对照组、传统玻璃化冷冻组和添加MitoQ的玻璃化冷冻组。对卵巢样本进行冷冻保存、解冻，并用苏木精和伊红（H&E）染色评估组织形态，用免疫荧光、透射电子显微镜和蛋白质免疫印迹分析评估线粒体变化。采用RNA测序（RNA-seq）探索潜在的保护机制。进行自体移植实验，通过H&E染色计数卵泡数量以及用酶联免疫吸附测定法（ELISA）测量血清雌二醇和抗苗勒管激素（AMH）水平来评估MitoQ对卵巢功能的长期影响。
发现1μM的MitoQ是玻璃化冷冻后维持卵泡形态的最佳浓度。它有效减少了线粒体氧化损伤，保留了线粒体形态，并调节了线粒体动力学蛋白（Drp1和Mfn2）的表达。RNA-seq和蛋白质免疫印迹分析表明，MitoQ抑制p38丝裂原活化蛋白激酶（MAPK）通路，从而减少细胞凋亡。此外，自体移植实验表明，与传统玻璃化冷冻相比，MitoQ处理显著增加了卵泡数量、雌二醇（E2）和抗苗勒管激素（AMH）水平。
MitoQ有效减轻玻璃化冷冻诱导的氧化损伤，维持线粒体稳态，并保留卵泡储备和内分泌功能。这些发现表明，MitoQ是卵巢组织冷冻保存中有价值的辅助剂，可显著改善癌症患者的生育力保存结果。
