Department of Obstetrics and Gynecology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea.
Department of Obstetrics and Gynecology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, South Korea.
Eur J Obstet Gynecol Reprod Biol. 2021 Jul;262:113-117. doi: 10.1016/j.ejogrb.2021.05.016. Epub 2021 May 11.
To investigate the influence of pretreatment of insulin on the phosphorylation of ERK1/2 by gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) in cultured human granulosa cells.
Human granulosa cells were collected from 20 women (age: 20-35 years) who underwent controlled ovarian hyperstimulation for in vitro fertilization and embryo transfer at Asan Medical Center (Seoul, South Korea). The presence of the receptors for insulin, GnRH, FSH, and LH in human granulosa cells was identified by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). The granulosa cells were treated with 10 nM insulin or 10 nM GnRH for 5 min or 30 min and with 10 nM FSH or 10 nM LH for 24 h or 48 h. The cells were also pretreated with insulin for 30 min prior to treatment with GnRH, FSH, or LH. Western blot analysis was used to analyze ERK1/2 phosphorylation.
RT-PCR showed that the receptors for insulin, GnRH, FSH, and LH were expressed in human granulosa cells. Insulin, GnRH, FSH, and LH could activate ERK1/2 phosphorylation. Pretreatment with insulin inhibited ERK1/2 phosphorylation induced by GnRH and FSH while augmenting ERK1/2 phosphorylation induced by LH.
Insulin might have a negative effect on GnRH and FSH regulation by attenuating the action of GnRH and FSH in the phosphorylation of ERK1/2 in human granulosa cells. In contrast, insulin might have a positive effect on LH regulation by potentiating the action of LH in the phosphorylation of ERK1/2. Our results showed that insulin is clearly an important regulator of human reproductive function at the ovarian level.
研究胰岛素预处理对促性腺激素释放激素（GnRH）、卵泡刺激素（FSH）和黄体生成素（LH）刺激人卵巢颗粒细胞 ERK1/2 磷酸化的影响。
本研究收集了 20 名在韩国首尔 Asan 医疗中心接受体外受精和胚胎移植控制性卵巢超排卵的妇女（年龄：20-35 岁）的人卵巢颗粒细胞。通过半定量逆转录聚合酶链反应（RT-PCR）鉴定人卵巢颗粒细胞中胰岛素、GnRH、FSH 和 LH 受体的存在。用 10 nM 胰岛素或 10 nM GnRH 处理颗粒细胞 5 分钟或 30 分钟，用 10 nM FSH 或 10 nM LH 处理 24 小时或 48 小时。用胰岛素预处理颗粒细胞 30 分钟，然后用 GnRH、FSH 或 LH 处理。用 Western blot 分析分析 ERK1/2 磷酸化。
RT-PCR 显示胰岛素、GnRH、FSH 和 LH 受体在人卵巢颗粒细胞中表达。胰岛素、GnRH、FSH 和 LH 均可激活 ERK1/2 磷酸化。胰岛素预处理抑制 GnRH 和 FSH 诱导的 ERK1/2 磷酸化，增强 LH 诱导的 ERK1/2 磷酸化。
胰岛素可能通过减弱 GnRH 和 FSH 在人卵巢颗粒细胞 ERK1/2 磷酸化中的作用，对 GnRH 和 FSH 调节产生负效应。相反，胰岛素可能通过增强 LH 在 ERK1/2 磷酸化中的作用对 LH 调节产生正效应。我们的研究结果表明，胰岛素在卵巢水平上显然是人类生殖功能的重要调节因子。
