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Abstract
Impaired ovarian follicle development, the hallmark of polycystic ovarian syndrome (PCOS), is believed to be due to the changes in expression of related genes such as follistatin (FST).
Expression of FST gene and methylation level of its promoter in theca cells from adult female rats, prenatally exposed to androgen excess, during different phases of the estrus cycle was determined and compared with controls.
Eight pregnant Wistar rats (experimental group) were treated by subcutaneous injection of 5 mg free testosterone on day 20 of pregnancy, while controls (n = 8) received 500 ml solvent. Based on observed vaginal smear, adult female offspring of mothers were divided into three groups. Levels of serum steroidogenic sexual hormones and gonadotropins, expression and promoter methylation of the FST gene were measured using ELISA, cyber-green real-time PCR and bisulfite sequence PCR (BSP), respectively.
Compared to controls, the relative expression of FST gene in the treated group decreased overall by 0.85 fold; despite significant changes in different phases, but no significant differences in methylation of FST promoter.
Our results reveal that manifestation of PCOS-like phenotype following prenatal exposure to excess androgen is associated with irregularity in expression of the FST gene during the estrus cycle.
卵泡发育障碍, 作为多囊卵巢综合征 (PCOS) 的特征, 被认为是由于相关基因如卵泡抑素 (FST) 表达的改变所致。
检测并与对照组比较成年雌性大鼠动情周期的不同时期中膜细胞内FST基因的表达, 及其启动子的甲基化水平、产前雄激素过多的暴露情况。
8只怀孕的Wistar大鼠 (实验组) 在怀孕第20天皮下注射5mg游离睾酮, 对照组 (n = 8) 接受500ml溶剂注射。根据观察到的阴道图片, 将这些大鼠的成年雌性后代被分为3组。血清类固醇性激素和促性腺激素水平、FST基因的表达和启动子甲基化分别采用ELISA、实时荧光PCR和亚硫酸氢钠测序PCR(BSP)检测。
与对照组相比, 治疗组FST基因的相对表达量降低0.85倍；尽管FST启动子的甲基化水平在不同时期有显著改变, 但无显著统计学差异。
我们的研究显示, 产前暴露于高雄激素环境后出现PCOS样的表现与动情周期中FST基因的不规律表达有关。
Acknowledgements
The authors wish to thank Ms. Niloofar Shiva for critical editing of English grammar and syntax of the manuscript.
Declaration of interest
The authors declare that there is no conflict of interest regarding the publication of this paper.