Guangdong Women and Children Hospital, Guangzhou Medical University, Guangzhou, China.
Guangzhou Municipal and Guangdong Provincial Key Laboratory of Protein Modification and Degradation, State Key Laboratory of Respiratory Disease, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, China.
Front Immunol. 2021 May 17;12:685724. doi: 10.3389/fimmu.2021.685724. eCollection 2021.
Hormones and immune imbalance are critical factors in polycystic ovary syndrome (PCOS). The alternation of immune microenvironment of oocytes may play a significant role in infertility of PCOS patients.
This study explores the role of follicular fluid microenvironment change in inflammatory pathways activation of granulosa cells (GCs) in PCOS women infertility.
We enrolled 27 PCOS patients and 30 controls aged 22 to 38 years who underwent IVF and collected their luteinized granulosa cells (LGCs). Meanwhile, a granulosa-like tumor cell line (KGN) as a cell-model assisted this study. Key inflammatory markers in human ovarian GCs and follicular fluid were detected by RT-qPCR, Western blotting, or ELISA. The KGN cells were treated with follicle supernatant mixed with normal medium to simulate the microenvironment of GCs in PCOS patients, and the inflammation indicators were observed. The assembly of NLRP3 inflammasomes was detected by immunofluorescence techniques. Dihydroethidium assay and EdU proliferation assay were used to detect ROS and cell proliferation by flow cytometry.
Compared with normal controls (n = 19), IL-1β (P = 0.0005) and IL-18 (P = 0.021) in the follicular fluid of PCOS patients (n = 20) were significantly increased. The NF-κB pathway was activated, and NLRP3 inflammasome was formatted in ovarian GCs of PCOS patients. We also found that inflammation of KGN cells was activated with LPS irritation or stimulated by follicular fluid from PCOS patients. Finally, we found that intracellular inflammation process damaged mitochondrial structure and function, which induced oxidative stress, affected cellular metabolism, and impaired cell proliferation.
Inflammatory microenvironment alteration in the follicular fluid of PCOS patients leads to activated inflammatory pathway in GCs, serving as a crucial factor that causes adverse symptoms in patients. This study provides a novel mechanism in the inflammatory process of PCOS.
激素和免疫失衡是多囊卵巢综合征（PCOS）的关键因素。卵母细胞免疫微环境的改变可能在 PCOS 患者不孕中起重要作用。
本研究探讨 PCOS 患者不孕中卵泡液微环境改变对颗粒细胞（GCs）炎症途径激活的作用。
我们招募了 27 名 PCOS 患者和 30 名年龄在 22 至 38 岁之间接受 IVF 的对照者，并收集了他们的黄体化颗粒细胞（LGCs）。同时，使用颗粒细胞样肿瘤细胞系（KGN）作为细胞模型辅助这项研究。通过 RT-qPCR、Western blot 或 ELISA 检测人卵巢 GCs 和卵泡液中的关键炎症标志物。用混合正常培养基的卵泡上清液处理 KGN 细胞，模拟 PCOS 患者 GCs 的微环境，观察炎症指标。通过免疫荧光技术检测 NLRP3 炎性小体的组装。用二氢乙啶测定法和 EdU 增殖测定法通过流式细胞术检测 ROS 和细胞增殖。
与正常对照组（n = 19）相比，PCOS 患者（n = 20）的卵泡液中白细胞介素-1β（IL-1β）（P = 0.0005）和白细胞介素-18（IL-18）（P = 0.021）显著增加。NF-κB 途径被激活，NLRP3 炎性小体在 PCOS 患者的卵巢 GCs 中形成。我们还发现，用 LPS 刺激或用 PCOS 患者的卵泡液刺激 KGN 细胞，会激活炎症。最后，我们发现细胞内炎症过程破坏了线粒体结构和功能，导致氧化应激，影响细胞代谢，损害细胞增殖。
PCOS 患者卵泡液中炎症微环境的改变导致 GCs 中炎症途径的激活，是导致患者不良症状的关键因素。本研究为 PCOS 炎症过程提供了一个新的机制。
