Effect of Platelet-rich Plasma on miR-۱۸۱a-۱, miR-۲۱, and miR-۱۲۲ Expression in Cryopreserved-Thawed Human Sperm: A Real-time PCR Study

Background and Aim: This study aimed to investigate the effect of platelet-rich plasma (PRP) on the expression of microRNA (miR)-۱۸۱a-۱, miR-۲۱, and miR-۱۲۲ in cryopreserved-thawed human sperm using quantitative real-time polymerase chain reaction (qRT-PCR). PRP, derived from autologous blood, is rich in growth factors and has shown potential for improving post-thaw sperm quality. Materials and Methods: RNA was extracted from fresh and cryopreserved-thawed sperm samples supplemented with varying concentrations of PRP (۰%, ۲.۵%, ۵%, ۱۰%, ۲۰%, and ۲۵%). Complementary DNA (cDNA) synthesis was performed using a stem-loop RT primer method, followed by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. Gene expression levels were normalized to U۶ small nuclear RNA and analyzed using the ۲−ΔΔCt method. Results: miR-۱۸۱a-۱ expression decreased across all PRP concentrations, but changes were not statistically significant (P>۰.۰۵). miR-۲۱ expression was significantly downregulated in ۱۰% (P=۰.۰۴۱) and ۲۵% (P<۰.۰۰۰۱) PRP samples. miR-۱۲۲ expression was significantly increased in the absence of PRP (P=۰.۰۰۸), but this effect was attenuated at higher PRP concentrations. Conclusion: PRP modulates miRNA expression in cryopreserved-thawed sperm in a concentration-dependent manner, suggesting a potential role in regulating post-thaw sperm integrity. These findings support further exploration of PRP as a cryoprotective agent in reproductive medicine.

کلیدواژه ها:

Frozen - thawed sperm ، miR-۱۸۱a-۱ ، miR-۲۱ ، miR-۱۲۲ ، Platelet-rich plasma (PRP) ، MicroRNA ، Real-time PCR