{"id":4,"title":"Optimized Proteomic and N-Glycoproteomic Profiling Reveals Aging-Associated Molecular Remodeling in Granulosa Cells","title_zh":"优化蛋白质组与N-糖蛋白组分析揭示颗粒细胞中衰老相关分子重塑","url":"https://pubmed.ncbi.nlm.nih.gov/42992916","doi":"10.1021/acs.jproteome.6c00614","pmid":"42992916","journal":"J Proteome Res","kind":"paper","lang":"en","published_at":"2026-11-06T00:00:00+00:00","fetched_at":"2026-11-08T03:43:49+00:00","authors":["Yan Y","Fan J","Yuan S","Wu Z","Wang J","Lu H"],"primary_topic":"oocyte","topics":[{"slug":"oocyte","score":7.0,"hits":["follicular","granulosa","oocyte","ovarian"]},{"slug":"omics-methods","score":5.0,"hits":["proteome","proteomic"]},{"slug":"repro-aging","score":1.0,"hits":["ovarian aging"]}],"topic_score":7.0,"preference_score":20.0,"preference_labels":["早期胚胎/卵母细胞蛋白质组"],"llm_score":76.0,"final_score":96.0,"score_dims":{"novelty":70,"rigor":68,"impact":72,"relevance":88,"translation":70},"recommend_reason":"该研究针对临床颗粒细胞样本量少、膜/分泌蛋白提取难的问题，系统优化并建立可复用的蛋白质组与N-糖蛋白组流程，同时提供年轻与高龄女性颗粒细胞差异图谱，对卵巢衰老研究和ART实验室多组学应用有参考价值；但样本量较小、缺乏独立验证，结论仍需谨慎解读。","keywords":["颗粒细胞","N-糖蛋白组","蛋白质组学","卵巢衰老","样本前处理","辅助生殖"],"selected":1,"story_id":43,"llm_provider":"deepseek","llm_model":"deepseek-flash","llm_effort":"low","source_key":"pubmed-embryo-omics","source_name":"PubMed · 胚胎/卵母细胞组学","source_tier":1,"mention_count":3,"has_interpretation":false,"journal_key":null,"journal_tier":"unknown","journal_if":null,"is_mock":false,"summary_zh":"本研究系统比较SDS、SDC和尿素三种蛋白提取流程及胰蛋白酶与LysC-胰蛋白酶消化策略，用于颗粒细胞蛋白质组和N-糖蛋白组分析，发现SDS流程整体表现最优，并将其应用于6例年轻与6例高龄女性的临床颗粒细胞样本，共鉴定7599个蛋白和4703个N-糖肽。整合分析发现12个差异表达蛋白和111个差异丰度N-糖肽，提示卵巢衰老伴随蛋白表达与N-糖基化改变，且糖基化变化可独立于蛋白丰度。该研究建立了临床颗粒细胞多组学分析的稳健流程，并为卵巢衰老机制提供新线索。","summary":"Granulosa cells (GCs) are essential for maintaining the follicular microenvironment and supporting oocyte development. However, the comprehensive profiling of the granulosa cell proteome and N-glycoproteome remains insufficiently characterized. Furthermore, the limited availability of clinical granulosa cell samples and the inherent difficulty in extracting membrane and secreted proteins necessitate a standardized sample preparation protocol. In this study, we systematically evaluated three protein extraction workflows (sodium dodecyl sulfate (SDS), sodium deoxycholate (SDC) and urea) alongside different digestion strategies (trypsin and tandem LysC-trypsin) for granulosa cell proteomic and N-glycoproteomic analyses. The SDS-based workflow showed superior overall performance and was applied to clinical granulosa cell samples from young (n = 6) and aged (n = 6) women. We successfully identified 7599 proteins and 4703 N-glycopeptides from the granulosa cell samples. Integrated proteomic and N-glycoproteomic analyses identified 12 differentially expressed proteins and 111 differentially abundant N-glycopeptides between young and aged GCs, suggesting alterations in protein expression and N-glycosylation associated with ovarian aging. Notably, changes in N-glycosylation were observed independently of changes in protein abundance. This study establishes a robust workflow for comprehensive proteomic and N-glycoproteomic profiling of clinical granulosa cell samples and provides novel insights into ovarian aging.","interpretation_basis":null,"fulltext_status":"not_found","interpretation":null,"fulltext":{"available":false,"status":"not_found","reason":"Europe PMC 无此记录：DOI 10.1021/acs.jproteome.6c00614","pmcid":null,"license":null,"chars":0,"source":"Europe PMC","fetched_at":"2026-11-08T03:59:10+00:00"}}