为研究多囊卵巢综合征(PCOS)患者卵巢颗粒细胞中c-fos 及CYP17 基因表达的变化,探讨c-fos 及CYP17 基因对PCOS 卵巢颗粒细胞雄激素分泌的影响及可能的作用机制,对拟行IVF/ICSI-ET 的PCOS 患者及非PCOS 患者分为对照组(非PCOS 患者)和PCOS 组(PCOS 患者),两组患者的卵巢颗粒细胞进行体外培养48 h,利用放射免疫法检测细胞上清液中的雌二醇(E2)、孕酮(P)、睾酮(T)的水平;利用2-NBDG 标记葡萄糖检测颗粒细胞的葡萄糖摄取能力;采用免疫蛋白印迹(WesternBlot)法分别评估颗粒细胞中c-fos 及CYP17 基因的表达变化。结果发现,PCOS 组患者与对照组患者相比,颗粒细胞培养上清液中睾酮水平升高(P<0.05)、孕酮水平降低(P<0.05)、雌二醇水平无明显变化(P>0.05),且颗粒细胞对2-NBDG 标记葡萄糖摄取明显降低(P<0.05),提示PCOS 患者卵巢颗粒细胞雄激素分泌异常增高,并存在糖代谢异常。与对照组患者相比,PCOS组患者颗粒细胞CYP17 基因表达升高(P<0.05),c-fos 基因表达降低(P<0.05),提示颗粒细胞c-fos 基因异常低表达,可能降低了对CYP17 基因的抑制作用,使CYP17 高表达,可能是卵巢颗粒细胞雄激素分泌增加的原因之一。
The objective of this study is to confirm the role of c-fos in the regulation of 17-alpha hydroxylase (CYP17) and androgen production in granulosa cells from Polycystic ovary syndrome (PCOS) patients. The ovarian granulosa cells from the PCOS and non- PCOS patients during IVF/ICSI–ET are isolated and divided into two groups: The PCOS group and the control group,then they are cultured in vitro for 48 h. The following techniques are then used: The radioimmunoassay to determine the level of estradiol, progesterone,testosterone; The 2-NBDG as the fluorescence probe to detect the glucose uptake in granulosa cells; And Western Blot technology to analyze the cellular expressions of c-fos, and CYP17. The results show that compared with the control group,the testosterone secretion within ovarian granulosa cells from PCOS patients is increased (P<0.05), the progesterone level is significantly lower (P<0.05), the estradiol (P>0.05) does not change significantly, and the 2-NBDG uptake in granulosa cells is lower (P<0.05), indicating that the testosterone secretion within ovarian granulosa cells from PCOS patients is significantly increased, and the glucose metabolic disorders are present in granulosa cells from PCOS patients. The Western Blot shows that CYP17 (P<0.05) has a high expression in ovarian granulosa cells from PCOS patients compared with the control group patients, and the expression of c-fos is decreased (P<0.05), indicating that the c-fos may be one of the factors responsible for the CYP17 repression, and it may be one of the reasons for the increase in the testosterone production of granulose cells.
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