Yin Qianqian, Zheng Jianhua, Cao Yijuan, Yan Xiaonan, Zhang Hong
Center for Reproductive Medicine, XuZhou Central Hospital, Xuzhou, Jiangsu, China.
Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Front Endocrinol (Lausanne). 2025 May 15;16:1579469. doi: 10.3389/fendo.2025.1579469. eCollection 2025.
Proteomic analysis was conducted on human follicular fluid (FF) using the 4D label-free method to identify proteins potentially influencing oocyte quality in hyperandrogenic (HA) polycystic ovary syndrome (PCOS) patients.
FF was collected from 3 different groups: HA PCOS patients, non-hyperandrogenic (NHA) PCOS patients, and controls. Protein profiles of FF from HA PCOS patients (n = 10) were constructed utilizing 4D label-free proteomics technology. Differentially expressed proteins were identified by comparing these profiles with those from NHA PCOS (n = 10) and control patients (n = 10). In addition, FF was collected from 34 HA, 33 NHA, and 23 control patients for enzyme-linked immunosorbent assay (ELISA) validation of differentially expressed proteins. Associations between the levels of differentially expressed proteins in FF and various embryonic outcome indicators were evaluated.
The HA group demonstrated significantly reduced normal cleavage rates, D3 available embryo rates, D3 high-quality embryo rates, available blastocyst rates, and high-quality blastocyst rates compared to the NHA and control groups (HA vs. NHA vs. Control, 88.3 vs. 93.6 vs. 94.23, 70.57 vs. 81.76 vs. 83.77, 42.49 vs. 56.39 vs. 61.83, 55.0 vs. 65.96 vs. 67.26, 27.62 vs. 45.19 vs. 44.75, respectively), with statistically significant differences ( < 0.05). 23 differentially expressed proteins were identified in FF profiles of the HA group relative to the control group, while 9 differentially expressed proteins were noted in comparison with the NHA group. Cross-comparison highlighted three potential target proteins: insulin-like growth factor binding protein 5 (IGFBP5), lysosomal-associated membrane protein 2 (LAMP2), and cadherin-5 (CDH5). Adjusting for age and body mass index (BMI), IGFBP5 levels in FF exhibited negative correlations with normal cleavage rate, D3 high-quality embryo rate, available blastocyst rate, and high-quality blastocyst rate ( 0.05). Similarly, LAMP2 levels were negatively correlated with normal cleavage rate, D3 available embryo rate, D3 high-quality embryo rate, and high-quality blastocyst rate ( < 0.05). CDH5 levels demonstrated positive correlations with D3 high-quality embryo rate and high-quality blastocyst rate ( < 0.05).
The proteins IGFBP5, LAMP2, and CDH5 may contribute to the mechanisms underlying the adverse effects of hyperandrogenism on oocyte quality in PCOS patients.
采用4D无标记方法对人卵泡液（FF）进行蛋白质组学分析，以鉴定可能影响高雄激素（HA）多囊卵巢综合征（PCOS）患者卵母细胞质量的蛋白质。
从3个不同组收集FF：HA PCOS患者、非高雄激素（NHA）PCOS患者和对照组。利用4D无标记蛋白质组学技术构建HA PCOS患者（n = 10）的FF蛋白质谱。通过将这些谱与NHA PCOS患者（n = 10）和对照患者（n = 10）的谱进行比较，鉴定差异表达的蛋白质。此外，从34例HA、33例NHA和23例对照患者中收集FF，用于差异表达蛋白质的酶联免疫吸附测定（ELISA）验证。评估FF中差异表达蛋白质水平与各种胚胎结局指标之间的关联。
与NHA组和对照组相比，HA组的正常分裂率、D3可用胚胎率、D3高质量胚胎率、可用囊胚率和高质量囊胚率显著降低（HA组 vs. NHA组 vs. 对照组，分别为88.3 vs. 93.6 vs. 94.23、70.57 vs. 81.76 vs. 83.77、42.49 vs. 56.39 vs. 61.83、55.0 vs. 65.96 vs. 67.26、27.62 vs. 45.19 vs. 44.75），差异具有统计学意义（<0.05）。相对于对照组，在HA组的FF谱中鉴定出23种差异表达蛋白质，与NHA组相比，有9种差异表达蛋白质。交叉比较突出了三种潜在的靶蛋白：胰岛素样生长因子结合蛋白5（IGFBP5）、溶酶体相关膜蛋白2（LAMP2）和钙黏蛋白-5（CDH5）。调整年龄和体重指数（BMI）后，FF中IGFBP5水平与正常分裂率、D3高质量胚胎率、可用囊胚率和高质量囊胚率呈负相关（<0.05）。同样，LAMP2水平与正常分裂率、D3可用胚胎率、D3高质量胚胎率和高质量囊胚率呈负相关（<0.05）。CDH5水平与D3高质量胚胎率和高质量囊胚率呈正相关（<0.05）。
蛋白质IGFBP5、LAMP2和CDH5可能参与了高雄激素血症对PCOS患者卵母细胞质量产生不利影响的潜在机制。
