Hedgehog pathway inhibition causes primary follicle atresia and decreases female germline stem cell proliferation capacity or stemness
Hedgehog pathway inhibition causes primary follicle atresia and decreases female germline stem cell proliferation capacity or stemness
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Hedgehog 通路抑制导致初级卵泡闭锁并降低雌性生殖干细胞增殖能力或干性
DOI：
10.1186/s13287-019-1299-5
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发表时间：
2019-07-05
影响因子：
7.5
通讯作者：
Pan, Zezheng
中科院分区：
文献类型：
--
作者：
Jiang, Yu;Zhu, Dantian;Pan, Zezheng
研究背景卵泡耗竭是引起卵巢早衰（premature ovarian failure，POF）和原发性卵巢功能不全（primary ovarian insufficiency，POI）的原因之一。因此，维持一定数量的雌性生殖干细胞（FGSC）是产生卵母细胞和补充原始卵泡池的最佳选择。调控FGSC增殖或干细胞的机制可能有助于恢复卵巢功能，但在出生后哺乳动物卵巢中仍不清楚。本研究旨在探讨抑制Hedgehog（Hh）信号通路的活性调节卵泡发育和FGSC增殖的机制。方法和结果为了了解Hh信号通路在卵巢衰老中的作用，我们在生理和病理小鼠中测定了不同生育年龄的Hh信号通路活性以及它们之间的相关性。此外，我们评估了卵泡的数量和发展和FGSC增殖或干细胞在体外和体内阻断Hh通路后的变化。此外，我们的目的是解释FGSC表型变化的机制之一，通过氧化应激和细胞凋亡与Hh通路特异性抑制剂GANT 61治疗诱导。结果显示，在生理老化和POF模型中，卵巢中Hh信号传导的活性降低，这与生殖系干细胞标志物Mvh和Oct 4的表达水平的趋势一致。在体外，阻断Hh通路会导致卵泡发育障碍，并在用GANT 61处理卵巢后耗尽卵巢生殖细胞和FGSC。当Hh活性被阻断时，培养的原代FGSC的增殖或干性降低。结果表明，GANT 61抑制FGSC细胞内抗氧化酶水平和Bcl-2/Bax比值降低，caspase 3表达水平升高，线粒体膜电位异常，ROS在该系统内积累。
BackgroundFollicle depletion is one of the causes of premature ovarian failure (POF) and primary ovarian insufficiency (POI). Hence, maintenance of a certain number of female germline stem cells (FGSCs) is optimal to produce oocytes and replenish the primordial follicle pool. The mechanism that regulates proliferation or stemness of FGSCs could contribute to restoring ovarian function, but it remains uncharacterized in postnatal mammalian ovaries. This study aims to investigate the mechanism by which inhibiting the activity of the hedgehog (Hh) signaling pathway regulates follicle development and FGSC proliferation.Methods and resultsTo understand the role of the Hh pathway in ovarian aging, we measured Hh signaling activity at different reproductive ages and the correlation between them in physiological and pathological mice. Furthermore, we evaluated the follicle number and development and the changes in FGSC proliferation or stemness after blocking the Hh pathway in vitro and in vivo. In addition, we aimed to explain one of the mechanisms for the FGSC phenotype changes induced by treatment with the Hh pathway-specific inhibitor GANT61 via oxidative stress and apoptosis. The results show that the activity of Hh signaling is decreased in the ovaries in physiological aging and POF models, which is consistent with the trend of expression levels of the germline stem cell markers Mvh and Oct4. In vitro, blocking the Hh pathway causes follicular developmental disorders and depletes ovarian germ cells and FGSCs after treating ovaries with GANT61. The proliferation or stemness of cultured primary FGSCs is reduced when Hh activity is blocked. Our results show that the antioxidative enzyme level and the ratio of Bcl-2/Bax decrease, the expression level of caspase 3 increases, the mitochondrial membrane potential is abnormal, and ROS accumulate in this system.ConclusionsWe observed that the inhibition of the Hh signaling pathway with GANT61 could reduce primordial follicle number and decrease FGSC reproductive capacity or stemness through oxidative damage and apoptosis.