摘要
目的:探讨miR-132在卵泡刺激素(FSH)介导的颗粒细胞孕酮合成中的作用。方法:FSH暴露处理原代颗粒细胞,利用实时定量RT-PCR检测miR-132的表达,放射免疫法测定P水平,用重组腺病毒感染颗粒细胞进行miR-132的过表达,Western Blot法检测Foxo3a的表达变化。结果:原代颗粒细胞经50ng/ml FSH暴露处理后,miR-132的表达水平显著上调,48h时表达量最高;感染Ad-miR132组的miR-132表达水平高于空白对照组和感染对照重组腺病毒组(P<0.001);感染Ad-miR132后颗粒细胞合成P水平高于空白对照组和感染对照重组腺病毒组(P<0.001);在FSH暴露处理后,Foxo3a表达呈现逐渐下降的趋势,提示miR-132对Foxo3a的表达有负调控作用。结论:miR-132促进FSH介导的颗粒细胞合成孕酮。
Objective: To explore the role of miR-132 in FSH mediated progesterone secretion in granulosa cells. Methods: Primary granulosa cells were exposed to FSH, the expression of miR-132 was detected by real-time RT-PCR, and the secretion level of progesterone was determined by radioimmunoassay (RIA). For the over expression of miR-132, Granulosa cells were infected by the recombinant adenovirus. Proteins were extracted for Western blot analysis to detect the expression of Foxo3a. Results:After the primary granulosa cells were exposed to 50ng/ml FSH, the expression of miR-132 was significantly up-regulated, and the highest expression occurred at 48 h. Compared with the cell in control recombinant adenovirus Ad-miRNC group and in blank control group, the expression level of cell in miR-132 infected by adenovirus Ad-miR132 group was significantly increased ( P <0.001), and the level of progesterone secreted by granulosa cells was significantly increased after infected by Ad-miR132 ( P <0.001). After FSH exposure, Foxo3a expression showed gradually decreasing, which suggested that miR-132 might negatively regulate Foxo3a expression. Conclusion: miR-132 promotes FSH mediated progesterone secretion of granulosa cells.
作者
郑盼盼
姚楠
高小博
骆海燕
邵馨妍
陆彩玲
ZHENG Panpan;YAO Nan;GAO Xiaobo;LUO Haiyan;SHAO Xinyan;LU Cailing(National Research Institute for Family Planning,Beijing,100081;Graduate School of Peking Union Medical College)
出处
《中国计划生育学杂志》
2019年第5期552-555,共4页
Chinese Journal of Family Planning
基金
中国医学科学院医学与健康科技创新工程项目资助(2018-I2M-1-004)
中央级公益性科研院所基本科研业务项目(2018GJM01)
