N-thiocarboxyanhydrides, amino acid-derived enzyme-activated H2S donors, enhance sperm mitochondrial activity in presence and absence of oxidative stress

Abstract

Background Hydrogen sulfide (H2S) donors are crucial tools not only for understanding the role of H2S in cellular function but also as promising therapeutic agents for oxidative stress-related diseases. This study aimed to explore the effect of amino acid-derived N-thiocarboxyanhydrides (NTAs), which release physiological H2S levels in the presence of carbonic anhydrase, on porcine sperm function during short-term incubation with and without induced oxidative stress. For this purpose, we employed two H2S-releasing NTAs with release half-lives (t1/2) in the range of hours that derived from the amino acids glycine (Gly-NTA) or leucine (Leu-NTA). Because carbonic anhydrase is crucial for H2S release from NTAs, we first measured the activity of this enzyme in the porcine ejaculate. Then, we tested the effect of Gly- and Leu-NTAs at 10 and 1 nM on sperm mitochondrial activity, plasma membrane integrity, acrosomal status, motility, motile subpopulations, and redox balance during short-term incubation at 38 °C with and without a reactive oxygen species (ROS)-generating system. Results Our results show that carbonic anhydrase is found both in spermatozoa and seminal plasma, with activity notably higher in the latter. Both Gly- and Leu-NTAs did not exert any noxious effects, but they enhanced sperm mitochondrial activity in the presence and absence of oxidative stress. Moreover, NTAs (except for Leu-NTA 10 nM) tended to preserve the sperm redox balance against the injuries provoked by oxidative stress, which provide further support to the antioxidant effect of H2S on sperm function. Both compounds also increased progressive motility over short-term incubation, which may translate into prolonged sperm survival. Conclusions The presence of carbonic anhydrase activity in mammalian spermatozoa makes NTAs promising molecules to investigate the role of H2S in sperm biology. For the first time, beneficial effects of NTAs on mitochondrial activity have been found in mammalian cells in the presence and absence of oxidative stress. NTAs are interesting compounds to investigate the role of H2S in sperm mitochondria-dependent events and to develop H2S-related therapeutic protocols against oxidative stress in assisted reproductive technologies.

Therapeutic Potential of Hydrogen Sulfide in Reproductive System Disorders

“…It was found that H 2 S-releasing agents based on amino acids simulated the release of physiological levels of H 2 S in the presence of carbonic anhydrase without exerting harmful effects on cell function. In addition, the H 2 S-releasing agents enhanced the forward motility of sperm during short-term treatment, which may prolong the survival of sperm [131]. These results suggest that the regulatory effects of H 2 S on sperm motility in vitro may contribute to assisted reproduction.…”

Antimicrobial peptides and proteins as alternative antibiotics for porcine semen preservation

“…A sperm aliquot (2 µL) was loaded into a pre-warmed Leja chamber (Leja Products BV, The Netherlands, chamber depth: 20 µm). The sperm motility and kinetic parameters were evaluated, as previously reported [26], using a Computer Assisted Sperm Analysis (CASA) (NIS-Elements, Nikon, Tokyo, Japan and Laboratory Imaging, Prague, Czech Republic), which consists of an Eclipse E600 tri-ocular phase contrast microscope (Nikon, Tokyo, Japan), equipped with a warming stage set at 38 ºC (Tokai Hit, Shizuoka, Japan), and a DMK 23UM021 digital camera (The Imaging Source, Bremen, Germany). The analysis was carried out using a 10× negative phase-contrast objective (Nikon, Tokyo, Japan).…”

“…The seminal ORP of the samples were determined as previously reported [26] with minor modifications. At the end of sperm incubation, a sample from each treatment was centrifuged at 16, 300 g for 5 minutes at room temperature.…”

Supplementation of sperm cryopreservation media with H2S donors enhances sperm quality, reduces oxidative stress, and improves in vitro fertilization outcomes

“…Considering the physiological and cytoprotective significance of H 2 S 49 , as well as the functional role of H 2 S donor compounds in biological systems 50 and also in sperm 22 , 29 , 58 to the best of our knowledge, in this study for the first time, we investigate the potential protective capabilities of fast-releasing (NaHS) and slow-releasing (GYY4137) H 2 S donors in maintenance of sperm functional parameters and in vitro fertilization rate in goat species following cryopreservation.…”