Objectives: Polycystic ovary syndrome (PCOS) is one of the most common endocrine diseases in women with reproductive age, which is associated with hyperandrogenism, insulin resistance, and ovulatory dysfunction. Progesterone receptor membrane component 1 (PGRMC1) can mediate progesterone to inhibit the apoptosis of ovarian granulosa cells and the growth of follicles, and to induce glucolipid metabolism disorder in ovarian granulosa cells, which is closely related to the occurrence and development of PCOS. This study aims to determine the expression of PGRMC1 in serum, ovarian tissue, ovarian granulosa cells, and follicular fluid in PCOS patients and non-PCOS patients, analyze the value of PGRMC1 in diagnosis and prognosis evaluation of PCOS, and investigate its molecular mechanism on ovarian granulosa cell apoptosis and glucolipid metabolism.
Methods: A total of 123 patients were collected from the Department of Obstetrics and Gynecology in Guangdong Women and Children Hospital (hereinafter referred to as "our hospital") from August 2021 to March 2022 and divided into 3 groups: a PCOS pre-treatment group (n=42), a PCOS treatment group (n=36), and a control group (n=45). The level of PGRMC1 in serum was detected by enzyme linked immunosorbent assay (ELISA). The diagnostic and prognostic value of PGRMC1 was evaluated in patients with PCOS by receiver operating characteristic (ROC) curve. Sixty patients who underwent a laparoscopic surgery from the Department of Obstetrics and Gynecology in our hospital from January 2014 to December 2016 were collected and divided into a PCOS group and a control group (n=30). The expression and distribution of PGRMC1 protein in ovarian tissues were detected by immunohistochemical staining. Twenty-two patients were collected from Reproductive Medicine Center in our hospital from December 2020 to March 2021, and they divided into a PCOS group and a control group (n=11). ELISA was used to detect the level of PGRMC1 in follicular fluid; real-time RT-PCR was used to detect the expression level of PGRMC1 mRNA in ovarian granulosa cells. Human ovarian granular cell line KGN cells were divided into a scrambled group which was transfected with small interfering RNA (siRNA) without interference and a siPGRMC1 group which was transfected with specific siRNA targeting PGRMC1. The apoptotic rate of KGN cells was detected by flow cytometry. The mRNA expression levels of PGRMC1, insulin receptor (INSR), glucose transporter 4 (GLUT4), very low density lipoprotein receptor (VLDLR), and low density lipoprotein receptor (LDLR) were determined by real-time RT-PCR.
Results: The serum level of PGRMC1 in the PCOS pre-treatment group was significantly higher than that in the control group (P<0.001), and the serum level of PGRMC1 in the PCOS treatment group was significantly lower than that in the PCOS pre-treatment group (P<0.001). The areas under curve (AUC) of PGRMC1 for the diagnosing and prognosis evaluation of PCOS were 0.923 and 0.893, respectively, and the cut-off values were 620.32 and 814.70 pg/mL, respectively. The positive staining was observed on both ovarian granulosa cells and ovarian stroma, which the staining was deepest in the ovarian granulosa cells. The average optical density of PGRMC1 in the PCOS group was significantly increased in ovarian tissue and ovarian granulosa cells than that in the control group (both P<0.05). Compared with the control group, the PGRMC1 expression levels in ovarian granulosa cells and follicular fluid in the PCOS group were significantly up-regulated (P<0.001 and P<0.01, respectively). Compared with the scrambled group, the apoptotic rate of ovarian granulosa cells was significantly increased in the siPGRMC1 group (P<0.01), the mRNA expression levels of PGRMC1 and INSR in the siPGRMC1 group were significantly down-regulated (P<0.001 and P<0.05, respectively), and the mRNA expression levels of GLUT4, VLDLR and LDLR were significantly up-regulated (all P<0.05).
Conclusions: Serum level of PGRMC1 is increased in PCOS patients, and decreased after standard treatment. PGRMC1 could be used as molecular marker for diagnosis and prognosis evaluation of PCOS. PGRMC1 mainly localizes in ovarian granulosa cells and might play a key role in regulating ovarian granulosa cell apoptosis and glycolipid metabolism.
目的: 多囊卵巢综合征(polycystic ovary syndrome，PCOS)是育龄妇女最常见的内分泌疾病之一，常伴有高雄激素血症、胰岛素抵抗及排卵功能障碍。孕激素受体膜组分1(progesterone receptor membrane component 1，PGRMC1)可介导孕酮抑制卵巢颗粒细胞凋亡及卵泡生长，诱导卵巢颗粒细胞葡萄糖及脂质代谢紊乱，与PCOS发生和发展密切相关。本研究旨在通过检测PGRMC1在PCOS患者及非PCOS患者血清、卵巢组织、卵泡液和卵巢颗粒细胞中的表达，分析PGRMC1对PCOS诊断及预后评估的价值，并探讨其调控卵巢颗粒细胞凋亡及糖脂代谢分子机制。方法: 纳入2021年8月至2022年3月就诊于广东省妇幼保健院(以下简称“我院”)妇产科门诊的患者123例，分为PCOS治疗前组(n=42)、PCOS治疗后组(n=36)、对照组(n=45)，采用酶联免疫吸附试验(enzyme linked immunosorbent assay，ELISA)检测其血清PGRMC1水平，并利用PGRMC1受试者操作特征(receiver operating characteristic，ROC)曲线判断PGRMC1对PCOS诊断、预后评估的临床价值。纳入2014年1月至2016年12月在我院妇产科行腹腔镜手术的患者60例，分为PCOS组和对照组(n=30)，采用免疫组织化学染色检测卵巢组织中PGRMC1蛋白质的表达及分布情况。纳入2020年12月至2021年3月就诊于我院生殖医学中心的患者22例，分为PCOS组和对照组(n=11)，采用ELISA检测卵泡液中PGRMC1水平；real-time RT-PCR检测卵巢颗粒细胞中PGRMC1 mRNA的表达水平。将人卵巢颗粒细胞系KGN细胞分为转染无干扰作用小干扰RNA(small interfering RNA，siRNA)的scrambled组和转染靶向抑制PGRMC1的特异性siRNA的siPGRMC1组，采用流式细胞术检测细胞凋亡率，real-time RT-PCR检测PGRMC1、胰岛素受体(insulin receptor，INSR)、葡萄糖转运蛋白4(glucose transporter 4，GLUT4)、极低密度脂蛋白受体(very low density lipoprotein receptor，VLDLR)、低密度脂蛋白受体(low density lipoprotein receptor，LDLR) mRNA的表达水平。结果: PCOS治疗前组血清PGRMC1水平显著高于对照组(P<0.001)，PCOS治疗后组血清PGRMC1水平显著低于PCOS治疗前组(P<0.001)；PGRMC1用于PCOS诊断和预后评估的曲线下面积(area under curve，AUC)分别为0.932和0.893，最佳截断值分别为620.32和814.70 pg/mL。PGRMC1在卵巢颗粒细胞和卵巢间质细胞上均有表达，并在颗粒细胞上染色最深，PCOS患者卵巢组织及颗粒细胞PGRMC1平均光密度值均明显高于对照组(均P<0.05)。与对照组相比，PCOS组PGRMC1在卵巢颗粒细胞及卵泡液中表达水平显著上调(分别为P<0.001及P<0.01)。与scrambled组相比，siPGRMC1组卵巢颗粒细胞凋亡率显著增加(P<0.01)，PGRMC1和INSR mRNA的表达水平均显著下调(分别为P<0.001和P<0.05)，GLUT4、VLDLR、LDLR mRNA表达水平均显著上调(均P<0.05)。结论: PCOS患者血清PGRMC1水平升高，并在规范治疗后下降，PGRMC1可作为PCOS诊断及预后评估的分子标志物。PGRMC1主要定位于卵巢颗粒细胞，可能在调控卵巢颗粒细胞凋亡和糖脂代谢中起关键作用。.
Keywords: glycolipid metabolism; ovarian granulosa cell; polycystic ovary syndrome; progesterone receptor membrane component 1.