DISRUPTION OF THE PEROXIREDOXIN SYSTEM AFFECTS HUMAN SPERM VIABILITY AND DNA INTEGRITY

MARÍA C. FERNÁNDEZ; ADEL R. MOAWAD; CRISTIAN O´FLAHERTY

Encuentro; Child Health Research Day; 2017

Background: Our laboratory is pioneer in characterizing the family of antioxidant called peroxiredoxins (PRDXs) and establishing their role in the antioxidant protection of human spermatozoa. Indeed, we found that spermatozoa from infertile men have inactive PRDXs. However, it is unknown whether human spermatozoa have an active PRDX re-activation system. Objective: We aimed to determine the impact of the inhibition of different players of the PRDX system on the reactive oxygen species (ROS) levels, viability and DNA oxidation in human spermatozoa.Materials and Methods: Highly motile spermatozoa from healthy donors were incubated with inhibitors of the PRDX system and viability, ROS levels and DNA oxidation were assessed by flow cytometry using Calcein-AM, MitoSOX-Red (O2-), APF and H2DCFDA (HO, ONOO-), and the anti-8-OHdG antibody, respectively.Results: We observed a significant decrease in viability in a dose dependence manner when incubated with inhibitors of the 2-Cys PRDXs (Conoidin A), calcium independent phospholipase A2 (PRDX6-PLA2) activity (MJ33) or the PRDX re-activation system (Auranofin, Ezatiostat, S-Hexylgluthatione and Ethacrynic acid) compared to controls (p