Ma Yabo, Ma Yuqin, Li Pengfei, Ma Fucheng, Yu Miao, Xu Jinrui, Yang Yi
Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western, Ningxia University, Yinchuan 750021, China; School of Life Sciences, Ningxia University, Yinchuan 750021, China.
Key Laboratory of Ministry of Education for Conservation and Utilization of Special Biological Resources in the Western, Ningxia University, Yinchuan 750021, China; School of Life Sciences, Ningxia University, Yinchuan 750021, China.
Cell Signal. 2025 Mar;127:111575. doi: 10.1016/j.cellsig.2024.111575. Epub 2024 Dec 20.
Polycystic ovary syndrome (PCOS) is a metabolic and endocrine disease that entails dysregulated ovulation, hyperandrogenism, and polycystic ovaries. While Wnt5a has been suggested to play key roles in follicular development and female fertility under normal conditions, its functions in the context of PCOS have yet to be established. This study was thus designed to explore the impact of Wnt5a on ovarian granulosa cell autophagy in PCOS, providing in vitro evidence in support of its role in this setting.
DHT-induced granulosa (KGN) cells were used as an in vitro model, and Wnt5a and autophagy-related protein levels in these cells were detected via Western blotting. Downregulating the expression of Wnt5a in KGN cells (by interference and inhibitor) was also performed, and Western blotting, RT-PCR, and immunofluorescence strategies were used to detect autophagy-related and PI3K/AKT/mTOR pathway-associated factors in this setting. In vivo, BOX5 was tested as a therapeutic inhibitor of Wnt5a in a murine model of DHEA-induced PCOS. Changes in ovarian morphology were detected through hematoxylin staining, while E2 and T hormone levels were quantified by ELISA, and autophagy-related factors in these animals were quantified through Western blotting, immunofluorescence, and immunohistochemistry.
Wnt5a and autophagy-related protein levels rose significantly in DHT-induced KGN cells. Following downregulation of the Wnt5a in these cells, a significant decrease in autophagy-related factor levels was noted relative to the DHT group, together with significant increases in pathway-related factors. In mice, BOX5 treatment was sufficient to restore serum levels of androgen and to improve polycystic ovarian changes, while also suppressing the levels of autophagy-associated factors within ovarian granulosa cells.
Wnt5a downregulation suppresses autophagy in PCOS granulosa cells through the activation of the PI3K/AKT/mTOR pathway, in addition to remediating polycystic ovarian changes and normalizing serum levels of sex hormones.
多囊卵巢综合征（PCOS）是一种代谢和内分泌疾病，伴有排卵失调、高雄激素血症和多囊卵巢。虽然已有研究表明Wnt5a在正常情况下对卵泡发育和女性生育能力起关键作用，但其在PCOS背景下的功能尚未明确。因此，本研究旨在探讨Wnt5a对PCOS患者卵巢颗粒细胞自噬的影响，为其在该环境中的作用提供体外证据。
以双氢睾酮（DHT）诱导的颗粒（KGN）细胞作为体外模型，通过蛋白质免疫印迹法检测这些细胞中Wnt5a和自噬相关蛋白的水平。还通过干扰和抑制剂下调KGN细胞中Wnt5a的表达，并采用蛋白质免疫印迹法、逆转录-聚合酶链反应（RT-PCR）和免疫荧光技术检测该环境下自噬相关因子以及磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（AKT）/哺乳动物雷帕霉素靶蛋白（mTOR）信号通路相关因子。在体内，将BOX5作为Wnt5a的治疗性抑制剂，应用于DHEA诱导的PCOS小鼠模型。通过苏木精染色检测卵巢形态变化，采用酶联免疫吸附测定（ELISA）法定量雌二醇（E2）和睾酮（T）激素水平，并通过蛋白质免疫印迹法、免疫荧光法和免疫组织化学法定量这些动物体内的自噬相关因子。
在DHT诱导的KGN细胞中，Wnt5a和自噬相关蛋白水平显著升高。下调这些细胞中Wnt5a的表达后，相对于DHT组，自噬相关因子水平显著降低，而信号通路相关因子水平显著升高。在小鼠中，BOX5治疗足以恢复雄激素的血清水平，改善多囊卵巢变化，同时还能抑制卵巢颗粒细胞内自噬相关因子的水平。
下调Wnt5a除了能改善多囊卵巢变化和使性激素血清水平正常化外，还能通过激活PI3K/AKT/mTOR信号通路抑制PCOS颗粒细胞中的自噬。
