Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, 2 Xueshi Road, Hangzhou, Zhejiang Province 310006, China.
Hum Reprod. 2010 Jun;25(6):1441-50. doi: 10.1093/humrep/deq078. Epub 2010 Apr 8.
The present study was designed to evaluate whether the alteration of aquaporin-9 (AQP-9) expression in granulosa cells (GCs) of patients with polycystic ovary syndrome (PCOS) was associated with the hyperandrogenism in follicular fluid (FF).
We recruited infertile women with PCOS (n = 14) and infertile women with tubal blockage (controls, n = 31) for this study. We examined total testosterone (TT), free androgen index (FAI), sex hormone-binding globulin (SHBG), FSH, LH and estradiol in FF. Real-time PCR and western blotting were performed to assess AQP-9 expression in GCs, including effects of dihydrotestosterone (DHT) in vitro.
AQP-9 protein was localized in the nucleus, cytoplasm and cell membrane of the human GCs. The TT, FAI and LH levels were all higher, and SHBG levels lower, in the FF of women with PCOS versus controls (P = 0.0145, 0.0001, 0.0191, 0.0001, respectively). AQP-9 mRNA level in GCs of patients with PCOS was tightly correlated with the TT, SHBG levels and FAI in FF (P = 0.0020, 0.0001, 0.0020, respectively). In vitro, DHT (10(-9) mol/l) decreased AQP-9 mRNA (lowest at 12 h) and protein levels in control GCs (P = 0.0005, 0.0247, respectively). The inhibitory effect of DHT on AQP-9 mRNA was attenuated by LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor (P = 0.0013). Fifty micromolar 4-(hydroxymercuri) benzoic acid sodium salt (PMB) and 10(-9) mol/l DHT blunted the swelling of GCs in hypotonic medium, respectively (P = 0.0350, 0.0027).
Hyperandrogenism in FF of women with PCOS inhibited AQP-9 in GCs through the PI3K pathway.
本研究旨在评估多囊卵巢综合征（PCOS）患者颗粒细胞（GCs）中水通道蛋白-9（AQP-9）表达的改变是否与卵泡液（FF）中的高雄激素血症有关。
我们招募了患有 PCOS 的不孕妇女（n=14）和患有输卵管阻塞的不孕妇女（对照组，n=31）进行这项研究。我们检测了 FF 中的总睾酮（TT）、游离雄激素指数（FAI）、性激素结合球蛋白（SHBG）、卵泡刺激素（FSH）、黄体生成素（LH）和雌二醇。实时 PCR 和 Western blot 用于评估 GCs 中的 AQP-9 表达，包括体外二氢睾酮（DHT）的作用。
AQP-9 蛋白定位于人 GCs 的核、细胞质和细胞膜。与对照组相比，PCOS 患者的 FF 中 TT、FAI 和 LH 水平均升高，SHBG 水平降低（P=0.0145、0.0001、0.0191、0.0001，分别）。PCOS 患者 GCs 中的 AQP-9 mRNA 水平与 FF 中的 TT、SHBG 水平和 FAI 呈紧密相关（P=0.0020、0.0001、0.0020，分别）。体外，DHT（10^-9mol/L）降低了对照组 GCs 中的 AQP-9 mRNA（最低在 12 小时）和蛋白水平（P=0.0005、0.0247，分别）。PI3K 抑制剂 LY294002 减弱了 DHT 对 AQP-9 mRNA 的抑制作用（P=0.0013）。50μmol/L 4-（羟汞）苯甲酸纳（PMB）和 10^-9mol/L DHT 分别抑制了低渗介质中 GCs 的肿胀（P=0.0350、0.0027）。
PCOS 患者 FF 中的高雄激素血症通过 PI3K 途径抑制 GCs 中的 AQP-9。
