【摘要】 目的 分析正常女性仅在卵泡液中特异性表达的microRNAs(miRNAs)的功能及可能的代谢途径。方法 选取2017年9月至2018年3月于河南省人民医院生殖医学中心因单纯输卵管因素不孕就诊的患者30例，于取卵日当天收取第一管清亮无血液污染的卵泡液，提取、纯化卵泡液外泌体，作为实验组；同时于取卵日当天早晨抽取入组患者外周血，提取、纯化血清外泌体，作为对照组。提取纯化外泌体miRNA,采用miRNA芯片检测技术分析两组外泌体miRNAs的表达谱，筛选出卵泡液中特有的miRNAs,生物信息学技术分析卵泡液中特有的miRNAs的功能及可能参与的调控通路。结果 卵泡液特异性外泌体miRNAs的靶基因主要富集40种信号通路，其中一些信号通路与卵子发育密切相关，如MAPK信号通路、胰岛素信号通路、TGF-beta、PI3K-Akt信号通路等；且对MAPK、TGF-β信号的靶基因进行PPI网络互作分析，筛选出与MAPK信号通路关联度较高的靶基因有FGFR1、KRAS、BDNF、BRAF、EGFR、IGF1R,与TGF-beta信号通路关联度较高的靶基因有SMAD2、ACVR1、BVP4、SMAD4、TGFBR1。结论 本研究表明卵泡液特异性外泌体miRNAs可能在调控卵子成熟、颗粒细胞增殖凋亡以及类固醇激素形成等方面发挥至关重要的作用。更多还原

【Abstract】 Objective To analyze the function and possible metabolic pathways of microRNAs(miRNAs) specifically expressed in follicular fluid in normal women. Methods From September 2017 to March 2018,30 patients with infertility due to simple tubal factors were selected from the Reproductive Medicine Center of Henan Provincial People ’s Hospital. The first tube of clear and blood-free follicular fluid was collected on the day of oocyte retrieval, and the exosomes of follicular fluid were extracted and purified as the experimental group. Meanwhile, peripheral blood of the enrolled patients was obtained to extract and purify the exosome as control groups. And then, the extracted exosomal miRNA in the two groups were detected using miRNA microarray analysis, and the specific miRNAs in follicular fluid were screened out. Bioinformatics technology was used to analyze the functions of the specific miRNAs in follicular fluid and the possible regulatory pathways involved. Results The target genes of follicle-specific exosome miRNAs were mainly enriched in 40 signaling pathways, some of which were closely related to ovum development, such as MAPK signaling pathway, insulin signaling pathway, TGF-beta, PI3K-Akt signaling pathway, etc. PPI network interaction analysis was performed on the target genes of MAPK and TGF-β signal, and the target genes with high correlation with MAPK signal pathway were screened out as FGFR1, KRAS, BDNF, BRAF, EGFR and IGF1R. SMAD2, ACVR1, BVP4, SMAD4 and TGFBR1 were the target genes with high correlation with TGF-beta signaling pathway. Conclusions This study suggests that follicular fluid specific exosome miRNAs may play a crucial role in regulating ovum maturation, granulosa cell proliferation and apoptosis, and steroid hormone formation.更多还原