Reproductive Medicine Center, Northern Jiangsu People's Hospital, Yangzhou City, Jiangsu Province 225000, China.
Reproductive Medicine Center, Northern Jiangsu People's Hospital, Yangzhou City, Jiangsu Province 225000, China.
Steroids. 2024 Dec;212:109516. doi: 10.1016/j.steroids.2024.109516. Epub 2024 Sep 21.
Polycystic ovary syndrome (PCOS) is a prevalent endocrine disorder in women of childbearing age. The role of Sprouty RTK Signaling Antagonist 4 (SPRY4) in ovarian function in PCOS was investigated herein, focusing on its regulation of ERK1/2 phosphorylation. PCOS models were established in mice using dehydroepiandrosterone (DHEA). The expression levels of SPRY4 in ovarian tissues were analyzed through RT-qPCR and immunohistochemistry. SPRY4 knockdown was achieved via lentivirus, and its effects on endocrine function, ovarian morphology, oxidative stress, and ERK1/2 phosphorylation were evaluated. Afterwards, granulosa cells were isolated and treated with DHEA and ERK2 agonist tert-Butylhydroquinone. The impacts of ERK2 activation on the regulation of SPRY4 knockdown were assessed using ELISA, fluorescent probes, western blotting, and biochemical assays. SPRY4 knockdown normalized the estrous cycle, reduced serum levels of testosterone, anti-Müllerian hormone, and luteinizing hormone/follicle-stimulating hormone ratio, and improved ovarian morphology. Additionally, SPRY4 knockdown alleviated oxidative stress by decreasing reactive oxygen species and malondialdehyde levels while increasing superoxide dismutase activity. It also restored steroidogenic enzyme expression, which were disrupted by DHEA induction. In vitro, SPRY4 knockdown enhanced granulosa cell viability and reduced ERK1/2 phosphorylation, with tert-Butylhydroquinone reversing these effects and restoring oxidative stress and steroidogenesis disruptions. Together, SPRY4 modulates ERK1/2 phosphorylation to influence oxidative stress and steroidogenesis in PCOS. Targeting SPRY4 may provide novel therapeutic avenues for improving ovarian function and managing PCOS.
多囊卵巢综合征(PCOS)是育龄妇女中常见的内分泌疾病。本文研究了 Sprouty RTK 信号拮抗剂 4(SPRY4)在 PCOS 卵巢功能中的作用,重点研究其对 ERK1/2 磷酸化的调节。使用脱氢表雄酮(DHEA)在小鼠中建立 PCOS 模型。通过 RT-qPCR 和免疫组织化学分析卵巢组织中 SPRY4 的表达水平。通过慢病毒实现 SPRY4 敲低,并评估其对内分泌功能、卵巢形态、氧化应激和 ERK1/2 磷酸化的影响。随后,分离出颗粒细胞,并使用 DHEA 和 ERK2 激动剂 tert-Butylhydroquinone 进行处理。使用 ELISA、荧光探针、western blot 和生化测定评估 ERK2 激活对 SPRY4 敲低调节的影响。SPRY4 敲低使发情周期正常化,降低血清睾酮、抗苗勒管激素和黄体生成素/卵泡刺激素比值,并改善卵巢形态。此外,SPRY4 敲低通过降低活性氧和丙二醛水平,同时增加超氧化物歧化酶活性来减轻氧化应激。它还恢复了被 DHEA 诱导破坏的类固醇生成酶表达。在体外,SPRY4 敲低增强了颗粒细胞活力并降低了 ERK1/2 磷酸化,tert-Butylhydroquinone 逆转了这些效应并恢复了氧化应激和类固醇生成破坏。总之,SPRY4 调节 ERK1/2 磷酸化以影响 PCOS 中的氧化应激和类固醇生成。靶向 SPRY4 可能为改善卵巢功能和治疗 PCOS 提供新的治疗途径。