Proteomic analysis of human follicular fluid based on the 4D label free method to identify proteins that may affect oocyte quality in hyperandrogenic PCOS patients.

OBJECTIVE

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.

METHODS

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.

RESULTS

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).

CONCLUSION

The proteins IGFBP5, LAMP2, and CDH5 may contribute to the mechanisms underlying the adverse effects of hyperandrogenism on oocyte quality in PCOS patients.