Song Minxue, Ni Lili, Ma Zebing, Zhong Xin, Liu Yibing, Kuang Jilin, Li Ping
Department of Gynecology, The Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China.
Department of Orthopedics, The Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, Hunan, China.
Front Genet. 2025 Aug 4;16:1626992. doi: 10.3389/fgene.2025.1626992. eCollection 2025.
Diminished ovarian reserve (DOR) becomes more common with age, and hypoxia is a key cause of apoptosis in ovarian granulosa cells. This study investigated the genetic links between hypoxia and DOR.
The GSE87201 dataset for DOR was sourced from Gene Expression Omnibus database, normalized for common differentially expressed genes (Co-DEGs), and identified Hypoxia-related differentially expressed genes (HRDEGs) GeneCards; Receiver Operating Characteristic (ROC) curves evaluated HRDEGs' diagnostic value, and protein-protein interaction networks were visualized with STRING and Cytoscape. Enrichment analyses included Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and immune cell infiltration compared ovarian reserve groups. A granulosa cell injury model was created using 4-hydroperoxycyclophosphamide (4-HC), with Quantitative real-time PCR and Western blot measuring FANCI and KAT2A expression, and Cell Counting Kit-8 assays and flow cytometry assessing cell proliferation and apoptosis.
Twelve hypoxia-related genes were differentially expressed between low ovarian reserve (LOR) and high ovarian reserve (HOR), with 17 linked to DOR; eight pathways differed between LOR and HOR. Six hub genes (, , , , , ) were enriched in Fanconi anemia and HIF-1 pathways, affecting microtubules, spindle formation, and cytoskeleton dynamics during mitosis. Immune cell infiltration analysis showed significant differences, with , , and correlating with immune populations. The DOR group had increased and levels compared to Control (two of the several genes that were matched were randomly selected for validation), alongside reduced cell viability and increased apoptosis.
, , , , , and may be diagnostic biomarkers for DOR, providing novel insights for future research into the pathogenesis of hypoxia-induced DOR.
卵巢储备功能减退（DOR）随年龄增长愈发常见，缺氧是卵巢颗粒细胞凋亡的关键原因。本研究调查了缺氧与DOR之间的遗传联系。
从基因表达综合数据库获取DOR的GSE87201数据集，对常见差异表达基因（Co-DEGs）进行标准化，并鉴定缺氧相关差异表达基因（HRDEGs）；利用基因卡片；通过受试者工作特征（ROC）曲线评估HRDEGs的诊断价值，并用STRING和Cytoscape可视化蛋白质-蛋白质相互作用网络。富集分析包括基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路，并比较卵巢储备组的免疫细胞浸润情况。使用4-氢过氧环磷酰胺（4-HC）建立颗粒细胞损伤模型，采用定量实时PCR和蛋白质印迹法检测FANCI和KAT2A的表达，并用细胞计数试剂盒-8法和流式细胞术评估细胞增殖和凋亡。
低卵巢储备（LOR）和高卵巢储备（HOR）之间有12个缺氧相关基因差异表达，其中17个与DOR相关；LOR和HOR之间有8条通路不同。6个枢纽基因（、、、、、）在范可尼贫血和HIF-1通路中富集，影响有丝分裂期间的微管、纺锤体形成和细胞骨架动力学。免疫细胞浸润分析显示存在显著差异，其中、和与免疫群体相关。与对照组相比，DOR组的和水平升高（从匹配的几个基因中随机选择两个进行验证），同时细胞活力降低，凋亡增加。
、、、、和可能是DOR的诊断生物标志物，为未来研究缺氧诱导的DOR发病机制提供了新见解。
