The Interaction of NO and H(2)S in Boar Spermatozoa under Oxidative Stress

SIMPLE SUMMARY: The most recent experiments performed on somatic cells describe the interaction of nitric oxide (NO) and hydrogen sulfide (H(2)S) on various levels. In male gametes, these two gasotransmitters have been studied individually up until today. Both NO and H(2)S participate in crucial sperm structural and functional changes before and after ejaculation. Moreover, the two gasotransmitters can augment or mitigate the negative impact of oxidative stress, depending on the concentration. Oxidative stress is a concomitant condition to various male reproduction disorders. In this experiment, we investigated in vitro the simultaneous application of NO and H(2)S donors, which was compared to single-donor application (NO or H(2)S) at 100 nM concentrations in boar spermatozoa under oxidative stress. The evaluation of sperm qualitative traits revealed a positive effect of the combination of the two donors in DD treatment on progressive motility and plasma membrane integrity compared to the control sample under oxidative stress (CtrOX). The results of this experiment indicate that the combination of NO and H(2)S donors exceeds the effect of single-donor application under given conditions. In conclusion, our research indicates the importance of gasotransmitter interaction in male gametes. ABSTRACT: Various recent studies dedicated to the role of nitric oxide (NO) and hydrogen sulfide (H(2)S) in somatic cells provide evidence for an interaction of the two gasotransmitters. In the case of male gametes, only the action of a single donor of each gasotransmitter has been investigated up until today. It has been demonstrated that, at low concentrations, both gasotransmitters alone exert a positive effect on sperm quality parameters. Moreover, the activity of gaseous cellular messengers may be affected by the presence of oxidative stress, an underlying condition of several male reproductive disorders. In this study, we explored the effect of the combination of two donors SNP and NaHS (NO and H(2)S donors, respectively) on boar spermatozoa under oxidative stress. We applied NaHS, SNP, and their combination (DD) at 100 nM concentration in boar spermatozoa samples treated with Fe(2+)/ascorbate system. After 90 min of incubation at 38 °C, we have observed that progressive motility (PMot) and plasma membrane integrity (PMI) were improved (p < 0.05) in DD treatment compared to the Ctr sample under oxidative stress (CtrOX). Moreover, the PMot of DD treatment was higher (p < 0.05) than that of NaHS. Similar to NaHS, SNP treatment did not overcome the PMot and PMI of CtrOX. In conclusion, for the first time, we provide evidence that the combination of SNP and NaHS surmounts the effect of single-donor application in terms of PMot and PMI in porcine spermatozoa under oxidative stress.