Small extracellular vesicles derived from embryonic stem cells restore ovarian function of premature ovarian failure through PI3K/AKT signaling pathway
Small extracellular vesicles derived from embryonic stem cells restore ovarian function of premature ovarian failure through PI3K/AKT signaling pathway
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胚胎干细胞来源的小细胞外囊泡通过PI3K/AKT信号通路恢复卵巢早衰的卵巢功能
DOI：
10.1186/s13287-019-1508-2
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发表时间：
2020-01-03
影响因子：
7.5
通讯作者：
Wu, Qingkai
中科院分区：
文献类型：
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作者：
Liu, Mengyu;Qiu, Yu;Wu, Qingkai
研究背景卵巢早衰（POF）严重影响女性生殖内分泌功能，是导致不孕的重要原因。先前的研究已经证明，来源于干细胞的小细胞外囊泡（sEVs）在组织再生中起重要作用。本研究旨在探讨胚胎干细胞来源的sEVs（ESCs-sEVs）对卵巢损伤的治疗作用及其分子机制。然后，将ESCs-sEV静脉内移植到POF小鼠中。分别于治疗后1周和2周采集小鼠血浆，采用ELISA法检测抗苗勒管激素（AMH）、雌二醇（E-2）和促卵泡激素（FSH）水平。H & E染色观察卵巢及卵泡形态，TUNEL法检测颗粒细胞凋亡。在体外，EdU和CCK-8测试用于评估ESCs-sEV刺激的培养颗粒细胞的增殖。结果ESCs-sEVs移植后，血清性激素水平基本恢复正常。此外，卵泡数量显著增加，凋亡细胞数量减少。体外实验结果显示，ESCs-sEVs能显著提高颗粒细胞的增殖率，并能显著增加颗粒细胞磷酸化PI3K和AKT的表达。结论ESCs-sEVs可通过调节PI3K/AKT信号通路改善卵巢功能，为卵巢早衰的临床治疗提供了一种新的途径。
BackgroundPremature ovarian failure (POF) has a great impact on reproductive endocrine function in females, and it is an important cause of infertility. Previous studies have demonstrated that small extracellular vesicles (sEVs) derived from stem cells play an important role in tissue regeneration. This study aimed to investigate the therapeutic effect of sEVs derived from embryonic stem cells (ESCs-sEVs) on damaged ovaries and explore the underlying molecular mechanisms.MethodsMice POF models were established by injecting mice with cyclophosphamide and busulfan. Then, ESCs-sEVs were intravenously transplanted into POF mice. The plasma of mice was harvested at 1 and 2weeks after treatment to analyze the levels of anti-Mullerian hormone (AMH), estradiol (E-2), and follicle stimulating hormone (FSH) by ELISA. The morphology of ovaries and follicles was observed by H&E staining, and apoptosis of granulosa cells was detected by TUNEL. In vitro, EdU and CCK-8 tests were used to evaluate the proliferation of cultured granulosa cells stimulated by ESCs-sEVs. Western blotting was used to determine the expression of PI3K/AKT and apoptotic-related proteins.ResultsAfter transplantation of ESCs-sEVs, the levels of serum sex hormones recovered to normal levels. In addition, the number of follicles was significantly increased, and the number of apoptotic cells was decreased. The results in vitro revealed that ESCs-sEVs could significantly improve the proliferation rate of granulosa cells and increase the expression of phosphorylated PI3K and AKT. Meanwhile, the positive effect on proliferation and the negative effect on apoptosis observed in granulosa cells were obviously decreased when the PI3K/AKT signaling pathway was inhibited.ConclusionOur findings suggested that ESCs-sEVs could improve ovarian function by regulating the PI3K/AKT signaling pathway, which could provide a promising clinical therapy for POF.