磷酸肌醇3-激酶/Akt/哺乳动物雷帕霉素靶复合物1(PI3K/Akt/mTORC1)途径在原始卵泡的激活中起关键作用。然而,过度激活和原始卵泡丢失可导致卵巢功能障碍。α-可溶性N-乙基马来酰亚胺敏感因子附着蛋白(α-SNAP)蛋白与PI3K/Akt/mTORCl信号传导有关,提示其可能参与卵泡激活。因此,本研究旨在探讨α-SNAP在PI3K/Akt/mTORC1信号通路激活中的作用及其减轻顺铂对卵巢功能影响的能力,使用体外和体内模型。
■我们用靶向α-SNAP的小干扰RNA(siRNA)转染KGN人卵巢颗粒细胞(GCs),以研究α-SNAP抑制对GC增殖和凋亡的影响,以及PI3K/Akt/mTORC1途径的活性。在老鼠模型中,在用顺铂治疗之前通过腺相关病毒递送α-SNAPsiRNA以评估其对卵泡活化和卵巢功能的影响。不同生长阶段的卵泡计数,西方印迹,和免疫组化分析进行检测裂解的caspase-3,Ki67,α-SNAP的表达,和p-mTOR。此外,抗苗勒管激素(AMH)的血清浓度通过酶联免疫吸附试验测定.
■体外,α-SNAP消耗通过抑制PI3K/Akt/mTORC1通路阻止GC增殖,从而表明其在调节细胞生长中的作用。在体内,α-SNAP敲低通过抑制PI3K/Akt/mTORC1信号通路和部分恢复AMH水平减弱顺铂诱导的原始卵泡过度激活。此外,caspase-3、Ki67、α-SNAP、p-mTOR在不同的卵泡生长阶段变化,提示对化疗诱导的卵巢损伤有保护作用。
■抑制α-SNAP可能通过抑制PI3K/Akt/mTORC1通路减弱GC增殖,从而减轻顺铂诱导的原始卵泡过度激活和丢失。靶向α-SNAP可能是预防化疗引起的卵巢损伤的新策略。然而,这些结论值得反复测试,未来必须进一步阐明α-SNAP的机理基础。
UNASSIGNED: The phosphoinositide 3-kinase/Akt/mammalian target of rapamycin complex 1 (PI3K/Akt/mTORC1) pathway plays a crucial role in the activation of primordial follicles. However, excessive activation and the loss of primordial follicles can lead to ovarian dysfunction. The alpha-soluble N-ethylmaleimide sensitive factor attachment protein (α-SNAP) protein has been implicated in PI3K/Akt/mTORCl signaling, suggesting its potential involvement in follicle activation. Thus, this study aimed to explore the role of α-SNAP in the activation of the PI3K/Akt/mTORC1 signaling pathway and its ability to mitigate the effects of cisplatin on ovarian function, using both in vitro and in vivo models.
UNASSIGNED: We transfected KGN human ovarian granulosa cells (GCs) with small interfering RNA (siRNA) targeting α-SNAP to investigate the effects of α-SNAP inhibition on GC proliferation and apoptosis, as well as on the activity of the PI3K/Akt/mTORC1 pathway. In a mouse model, α-SNAP siRNA was delivered via an adeno-associated virus before treatment with cisplatin to assess its effects on follicle activation and ovarian function. Follicle counts at various growth stages, western blotting, and immunohistochemistry analyses were conducted to detect the expression of cleaved caspase-3, Ki67, α-SNAP, and p-mTOR. Additionally, the serum concentrations of anti-Müllerian hormone (AMH) were measured through an enzyme-linked immunosorbent assay.
UNASSIGNED: In vitro, α-SNAP depletion prevented GC proliferation by inhibiting the PI3K/Akt/mTORC1 pathway, thereby indicating its role in the regulation of cell growth. In vivo, α-SNAP knockdown attenuated the cisplatin-induced overactivation of primordial follicles by suppressing the PI3K/Akt/mTORC1 signaling pathway and partially restoring AMH levels. In addition, the expression and distribution patterns of cleaved caspase-3, Ki67, α-SNAP, and p-mTOR varied across different follicular growth stages, suggesting a protective effect against chemotherapy-induced ovarian damage.
UNASSIGNED: Inhibiting α-SNAP may attenuate GC proliferation by suppressing the PI3K/Akt/mTORC1 pathway, thereby mitigating the overactivation and loss of primordial follicles induced by cisplatin. Targeting α-SNAP may emerge as a novel strategy to prevent ovarian damage resulting from chemotherapy. However, these conclusions warrant repeated testing, and the mechanistic underpinnings of α-SNAP must be further elucidated in the future.