Xu Rongxiang Regenerative Medicine Research Center, Binzhou Medical University, Yantai, 264003, China.
Department of Histology and Embryology, College of Basic Medicine, Binzhou Medical University, Yantai, 264003, China.
Sci Rep. 2024 Aug 16;14(1):19008. doi: 10.1038/s41598-024-69381-9.
Premature ovarian insufficiency (POI), a major cause of female infertility, is defined as follicular atresia and a rapid loss of germ cells in women of reproductive age due to ovarian failure. Recently, findings from several studies have indicated that human umbilical cord mesenchymal stem cells (hUMSCs) can alleviate ovarian dysfunction resulting from POI. However, the mechanisms underlying this effect require further clarification. In this study, a mouse model of POI was established as achieved with an intraperitoneal injection of cyclophosphamide (CTX) into female C57BL/6J mice in vivo. These POI mice received a 1-week intervention of hUMACs. In addition, an in vitro POI model was also included. The cultured supernatants of hUMSCs and glycogen synthase kinase 3 beta (GSK3β) inhibitor (SB216763) were used to treat theca cells (TCs) exposed to CTX. Hematoxylin and Eosin (H&E) staining and Enzyme-linked immunosorbent assay (ELISA) were used to assess ovarian structure and morphology, as well as endocrine function in these POI mice. Based on results from the ELISA and JC-1 labeling, CTX exerted significant detrimental effects on testosterone levels and the mitochondrial membrane potential in TCs. Subsequently, Western Blot, Immunofluorescence staining (IF), and Quantitative real-time polymerase chain reaction (qRT-PCR) were used to evaluate various indicators of testosterone synthesis function and mitochondrial dynamics in ovaries and TCs of POI mice. In vivo, dysfunctions in ovarian structure and function in the POI mouse model were effectively restored following hUMSCs treatment, and abnormalities in hormone synthesis were significantly reduced. Furthermore, when the stem cell supernatants of hUMSCs were applied to TCs in vitro we found that GSK3β expression was reduced, the imbalance of mitochondrial dynamics was alleviated, and the ability of mitochondrial testosterone synthesis was increased. Taken together, our results indicate that hUMSCs treatment can restore the imbalance of mitochondrial dynamics and restart testosterone synthesis of TCs by suppressing GSK3β expression, ultimately alleviating POI damage.
卵巢早衰（POI）是女性不孕的主要原因之一，其定义为育龄妇女由于卵巢衰竭而导致卵泡闭锁和生殖细胞迅速丢失。最近，多项研究结果表明，人脐带间充质干细胞（hUMSC）可以缓解 POI 引起的卵巢功能障碍。然而，这种作用的机制尚需进一步阐明。在这项研究中，通过腹腔内注射环磷酰胺（CTX）建立了 POI 小鼠模型，该模型在体内作用于 C57BL/6J 雌性小鼠。这些 POI 小鼠接受了为期一周的 hUMAC 干预。此外，还包括体外 POI 模型。用 hUMSC 的培养上清液和糖原合成酶激酶 3β（GSK3β）抑制剂（SB216763）处理暴露于 CTX 的颗粒细胞（TCs）。用苏木精和伊红（H&E）染色和酶联免疫吸附试验（ELISA）评估这些 POI 小鼠的卵巢结构和形态以及内分泌功能。根据 ELISA 和 JC-1 标记的结果，CTX 对 TCs 中的睾酮水平和线粒体膜电位产生了显著的有害影响。随后，使用 Western Blot、免疫荧光染色（IF）和实时定量聚合酶链反应（qRT-PCR）评估 POI 小鼠卵巢和 TCs 中睾酮合成功能和线粒体动力学的各种指标。在体内，hUMSC 处理有效恢复了 POI 小鼠模型中卵巢结构和功能的功能障碍，并且显著减少了激素合成的异常。此外，当将 hUMSC 的干细胞上清液应用于体外 TCs 时，我们发现 GSK3β 表达减少，线粒体动力学失衡得到缓解，并且线粒体睾酮合成能力增加。总之，我们的结果表明，hUMSC 处理通过抑制 GSK3β 表达可以恢复 TCs 中线粒体动力学的失衡并重新启动睾酮合成，从而缓解 POI 损伤。
