Xu Bo, Deng Yu, Dai Wenjie, Shang Yitong, Ding Liyang, Zhang Zhen, Yang Hong, He Tiantian, Pei Xiuying, Fu Xufeng
Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, School of Basic Medical Science, Key Laboratory of Reproduction and Genetics of Ningxia Hui Autonomous Region, Ningxia Medical University, Yinchuan, 750004, China.
Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, School of Basic Medical Science, Key Laboratory of Reproduction and Genetics of Ningxia Hui Autonomous Region, Ningxia Medical University, Yinchuan, 750004, China.
Free Radic Biol Med. 2025 Aug 16;236:28-40. doi: 10.1016/j.freeradbiomed.2025.05.396. Epub 2025 May 16.
Oxidative stress-induced death of ovarian granulosa cells (GCs) is a major driver of ovarian functional disorders associated with follicular atresia. Ferroptosis is a key factor in the onset and progression of various ovarian oxidative stress-related diseases, making it a potential target for enhancing reproductive health. Recently, 3D cultured human umbilical cord mesenchymal stem cells (3D hUCMSCs) spheroids have exhibited promising advantages in protecting GCs from oxidative damage. However, it is unclear whether they represent a viable therapeutic strategy for mitigating reproductive failure associated with abnormal follicular atresia by modulating ferroptosis. This study demonstrated that 3D hUCMSC spheroids can effectively protect GCs from hydrogen peroxide (HO)-induced oxidative stress and ferroptosis. Additionally, iron overload and lipid peroxidation are two essential features of ferroptosis. 3D hUCMSC spheroids effectively regulate iron uptake and storage to mitigate HO-induced iron overload. Furthermore, 3D hUCMSC spheroids mitigate lipid peroxidation induced by HO by restoring GSH metabolic balance and preventing GPX4 inactivation. Mechanistically, the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway was significantly activated by 3D hUCMSC spheroid treatment. Our findings reveal that Nrf2 knockdown inhibited the 3D hUCMSC spheroids-mediated resistance of GCs to HO-induced ferroptosis, and Nrf2 knockdown led to increased iron uptake, resulting in substantial lipid peroxidation through the Fenton reaction, thereby making GCs more susceptible to ferroptosis. This process may involve the ROS-Nrf2-Fe cycle. Significantly, 3D hUCMSC spheroids can mitigate HO-induced ferroptosis in GCs by regulating the ROS-Nrf2-Fe cycle. Finally, we confirmed the above results that 3D hUCMSC spheroids ameliorate ovarian oxidative damage in premature ovarian failure (POF) rats. In conclusion, we demonstrated that 3D hUCMSC spheroids regulate oxidative stress and iron homeostasis through the Nrf2 pathway, thereby providing a potential therapeutic target for anovulatory disorders.
氧化应激诱导的卵巢颗粒细胞（GCs）死亡是与卵泡闭锁相关的卵巢功能障碍的主要驱动因素。铁死亡是各种卵巢氧化应激相关疾病发生和发展的关键因素，使其成为增强生殖健康的潜在靶点。最近，3D培养的人脐带间充质干细胞（3D hUCMSCs）球体在保护GCs免受氧化损伤方面显示出有前景的优势。然而，尚不清楚它们是否代表一种可行的治疗策略，通过调节铁死亡来减轻与异常卵泡闭锁相关的生殖功能衰竭。本研究表明，3D hUCMSC球体可以有效保护GCs免受过氧化氢（HO）诱导的氧化应激和铁死亡。此外，铁过载和脂质过氧化是铁死亡的两个基本特征。3D hUCMSC球体有效调节铁的摄取和储存，以减轻HO诱导的铁过载。此外，3D hUCMSC球体通过恢复谷胱甘肽（GSH）代谢平衡和防止谷胱甘肽过氧化物酶4（GPX4）失活来减轻HO诱导的脂质过氧化。机制上，3D hUCMSC球体处理显著激活了核因子红细胞2相关因子2（Nrf2）信号通路。我们的研究结果表明，Nrf2基因敲低抑制了3D hUCMSC球体介导的GCs对HO诱导的铁死亡的抗性，并且Nrf2基因敲低导致铁摄取增加，通过芬顿反应导致大量脂质过氧化，从而使GCs更容易发生铁死亡。这个过程可能涉及活性氧（ROS）-Nrf2-铁循环。值得注意的是，3D hUCMSC球体可以通过调节ROS-Nrf2-铁循环来减轻HO诱导的GCs铁死亡。最后，我们证实了上述结果，即3D hUCMSC球体改善了卵巢早衰（POF）大鼠的卵巢氧化损伤。总之，我们证明了3D hUCMSC球体通过Nrf2途径调节氧化应激和铁稳态，从而为无排卵性疾病提供了一个潜在的治疗靶点。
