年龄相关性生殖功能减退的卵泡液代谢组学研究

Study on the follicular fluid metabolomics of age-related reproductive dysfunction

目的:通过对不同年龄段体外受精(IVF)患者的卵泡液进行代谢组学研究,鉴定出代谢差异物,探究其与卵母细胞的关系,从卵泡液代谢组层面阐述年龄影响生殖功能的机制。方法:对191例IVF患者卵泡液进行代谢组学分析,21～27岁47例,28～34岁74例,35～41岁41例,42～48岁29例。采用超高液相色谱-高分辨质谱联用技术将卵泡液样品分别在正离子、负离子模式下进行采集,通过主成分分析法(PCA)、偏最小二乘法-判别分析法(PLS-DA)得到四组间差异性物质质荷比。根据得到的质量数、同位素丰度比得到与其相匹配的分子式,鉴定出差异物,把这些差异物通过KEGG聚类分析,指向差异通路,鉴定这些差异物质涉及的代谢通路和所指向的化合物。结果:随着年龄增加,卵母细胞质量及数量呈下降趋势,期间伴随卵泡液代谢物变化,差异物主要涉及氨基酸、脂质、激素、维生素等。下调物15种:6-氧代己酸酯,L-苯丙氨酸,脯氨酸,十六烷酸/棕榈酸,亚油酸,花生四烯酸,油酸,硫酸脱氢表雄酮,溶血性磷脂酰胆碱(16∶1),溶血性磷脂酰胆碱(20∶4),溶血性磷脂酰胆碱(20∶3),25-二羟维生素D3,5-去氢表甾醇,27-羟基胆固醇,5-胆甾烷-3α,7α,12α,23,25-戊醇;上调物3种:溶血性磷脂酰胆碱(18∶3),溶血性磷脂酰胆碱(18∶1),13,14-二氢视黄醇。结论:女性生殖能力随着年龄增加而减退,期间伴随卵泡液代谢物变化,从卵泡液代谢组学层面揭示了年龄影响生殖功能的机制。

Objective： Follicular fluid in patients with different ages IVF metabonomics studies,identified the metabolic differences,explore its relationship with the oocytes,age from the set of follicular fluid metabolism level in this paper,the influence mechanism of reproductive function.Methods： A total of 191 patients with IVF were enrolled in this study,including 21 years old≤group A≤27 years old,47 cases,28 ≤group B ≤34 years old,74 cases,35 years old≤group C≤41 years old,41 cases,42 years old ≤group D ≤48 years old,29 cases,using the ultra-high performance liquid chromatography-high resolution mass spectrometry combined with follicular fluid samples were collected in positive and negative ion mode,through Principal component analysis（ PCA） and partial least-squares-discriminant analysis（ PLS-DA） were used to get the mass-to-charge ratios of the four groups.According to the masses obtained,the isotopic abundance ratios were matched Molecular formula,identify the difference,and these differences through KEGG clustering analysis,pointing to the differential pathways,further identification of these differential substances involved in the metabolic pathways and pointing compounds.Results： The quality and quantity of oocyte were decreasing with age,along with the change of follicular fluid metabolites,during different material mainly relates to amino acids,lipids,hormones,vitamins,etc. There are 15 were down-regulated： 6-Oxohexanoate,L-Phenylalanine,Proline,Hexadecanoic acid,Linoleate,Arachidonate,Oleic acid,Docosahexaenoic acid,Lyso PC（ 16 ∶ 1）,Lyso PC（ 20 ∶ 4）,Lyso PC（ 20 ∶ 3）,25-Hydroxyvitamin D3,5-Dehydroepisterol,27-Hydroxycholesterol,5 beta-cholestane-3 alpha,7 alpha,12 alpha,23,25-pento,and three kinds of up-regulated substances： Lyso PC（ 18 ∶ 3）,Lyso PC（ 18 ∶ 1）,13,14-Dihydroretinol.Conclusions： The reproductive ability of females was decreased with age,and the changes of metabolites of follicular fluid were observed during the period,and the mechanism of reproductive success was revealed from the perspective of follicular fluid metabolomics.