Epicatechin Provides Antioxidant Protection to Bovine Spermatozoa Subjected to Induced Oxidative Stress

Epicatechin (EPI) is a natural flavonoid with antibacterial, anti-inflammatory and anti-cancer properties. Furthermore, the molecule exhibits powerful reactive oxygen species (ROS) scavenging and metal-chelating properties. In this study, we assessed the efficiency of EPI to reverse ROS-mediated alterations to the motility, viability, DNA integrity and oxidative profile of bovine spermatozoa. For the first experiment, spermatozoa were washed out of fresh semen and exposed to 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI and 100 μmol/L EPI in the presence of ferrous ascorbate (FeAA) during a 6 h in vitro culture. For the second experiment, the ejaculates were split into aliquots and cryopreserved with a commercial semen extender supplemented with 12.5 μmol/L EPI, 25 μmol/L EPI, 50 μmol/L EPI, 100 μmol/L EPI or containing no supplement. Sperm motility was assessed using the computer-aided sperm analysis and the cell viability was studied with the metabolic activity test. ROS production was quantified using luminometry, and DNA fragmentation was evaluated using the chromatin dispersion test. Cell lysates were prepared at the end of the culture in order to assess the concentration of protein carbonyls and malondialdehyde. Exposure to FeAA led to a significantly reduced sperm motility (p < 0.001), mitochondrial activity (p < 0.001), but increased the generation of ROS (p < 0.001), as well as oxidative damage to proteins (p < 0.001), DNA (p < 0.001) and lipids (p < 0.001). EPI supplementation, particularly at a concentration range of 50–100 μmol/L, resulted in higher preservation of the spermatozoa vitality (p < 0.001). Furthermore, 50–100 μmol/L EPI were significantly effective in the prevention of oxidative damage to sperm proteins (p < 0.001), lipids (p < 0.001) and DNA (p < 0.01 in relation to 50 μmol/L EPI; p < 0.001 with respect to 100 μmol/L EPI). In the case of the cryopreserved spermatozoa, the administration of 50–100 μmol/L EPI resulted in higher sperm motility (p < 0.001) and mitochondrial activity (p < 0.001). ROS production, the number of protein carbonyls, lipid peroxidation as well as oxidative DNA damage were found to be significantly decreased particularly in samples cryopreserved in the presence of 100 μmol/L EPI (p < 0.001). Our results suggest that EPI could behave as an effective antioxidant which may prevent oxidative insults to spermatozoa, and thus, preserve their vitality and functionality. Nevertheless, its potential to achieve higher fertilization rates in reproductive technologies needs to be validated.

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Lycopene and resveratrol improve post-thaw bull sperm parameters: sperm motility, mitochondrial activity and DNA integrity

Andrologia ◽

10.1111/and.12301 ◽

2014 ◽

Vol 47 (5) ◽

pp. 545-552 ◽

Cited By ~ 40

Author(s):

M. N. Bucak ◽

M. B. Ataman ◽

N. Başpınar ◽

O. Uysal ◽

M. Taşpınar ◽

...

Keyword(s):

Sperm Motility ◽

Sperm Parameters ◽

Mitochondrial Activity ◽

Dna Integrity ◽

Bull Sperm

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Structure-based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies

Scientific Reports ◽

10.1038/s41598-021-91636-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Mutyala Satish ◽

Sandhya Kumari ◽

Waghela Deeksha ◽

Suman Abhishek ◽

Kulhar Nitin ◽

...

Keyword(s):

Sperm Motility ◽

In Silico ◽

Assisted Reproductive Technologies ◽

Ex Vivo ◽

Binding Free Energy ◽

Reproductive Technologies ◽

Early Embryo ◽

Dna Integrity ◽

Binding Studies

AbstractPhosphodiesterase (PDE) inhibitors, such as pentoxifylline (PTX), are used as pharmacological agents to enhance sperm motility in assisted reproductive technology (ART), mainly to aid the selection of viable sperm in asthenozoospermic ejaculates and testicular spermatozoa, prior to intracytoplasmic sperm injection (ICSI). However, PTX is reported to induce premature acrosome reaction (AR) and, exert toxic effects on oocyte function and early embryo development. Additionally, in vitro binding studies as well as computational binding free energy (ΔGbind) suggest that PTX exhibits weak binding to sperm PDEs, indicating room for improvement. Aiming to reduce the adverse effects and to enhance the sperm motility, we designed and studied PTX analogues. Using structure-guided in silico approach and by considering the physico-chemical properties of the binding pocket of the PDEs, designed analogues of PTX. In silico assessments indicated that PTX analogues bind more tightly to PDEs and form stable complexes. Particularly, ex vivo evaluation of sperm treated with one of the PTX analogues (PTXm-1), showed comparable beneficial effect at much lower concentration—slower AR, higher DNA integrity and extended longevity of spermatozoa and superior embryo quality. PTXm-1 is proposed to be a better pharmacological agent for ART than PTX for sperm function enhancement.

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Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure

Biology ◽

10.3390/biology9090254 ◽

2020 ◽

Vol 9 (9) ◽

pp. 254 ◽

Cited By ~ 3

Author(s):

Jaime Catalán ◽

Marion Papas ◽

Sabrina Gacem ◽

Yentel Mateo-Otero ◽

Joan E. Rodríguez-Gil ◽

...

Keyword(s):

Sperm Motility ◽

Consumption Rate ◽

Red Light ◽

Light Irradiation ◽

Mitochondrial Activity ◽

Dna Integrity ◽

O2 Consumption ◽

Sperm Viability ◽

Intracellular Atp ◽

Atp Levels

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

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In Vitro Studying the Effect of Adding Autologous Platelet Rich Plasma (PRP) to the Human Semen on the Sperm DNA Integrity

Iraqi Journal of Embryos and Infertility Researches ◽

10.28969/ijeir.v10.i2.r7 ◽

2021 ◽

Vol 10 (2) ◽

pp. 90-100

Author(s):

Dhafer Hamdan ◽

Ali Rahim ◽

Ula Al-Kawaz

Keyword(s):

Dna Fragmentation ◽

Assisted Reproductive Technologies ◽

Platelet Rich Plasma ◽

Reproductive Technologies ◽

Human Reproduction ◽

Sperm Chromatin ◽

Dna Integrity ◽

Sperm Dna Integrity ◽

Sperm Dna ◽

Dispersion Test

For conception and the development of healthy embryos, sperm DNA integrity is crucial. According to a growing body of studies, there is a strong correlation between sperm DNA damage and male infertility. Among the new medicines being developed in the medical field, the application of Platelet Rich Plasma (PRP) in human reproduction has yet to be examined. A total of 100 semen samples were used in the current experimental investigation. From November 2020 to June 2021, the research was conducted at the High Institute for Infertility Diagnosis and Assisted Reproductive Technologies. Masturbation was used to get an ejaculated semen sample. After semen analysis, the samples were separated into two equal parts, one without autologous PRP and the other with 2% autologous PRP, with the DNA fragmentation assessed using the Sperm Chromatin Dispersion Test. There was highly significant reduction in DNA fragmentation index (p < 0.001). The mean sperm DNA integrity was reduced after adding PRP (33.85±16.73 vs 38.55±16.64), Mean (± SE). PRP has been shown to improve human sperm DNA integrity.

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Curcumin offers antioxidant protection to cryopreserved bovine semen

Czech Journal of Animal Science ◽

10.17221/33/2017-cjas ◽

2018 ◽

Vol 63 (No. 7) ◽

pp. 247-255 ◽

Cited By ~ 3

Author(s):

E. Tvrdá ◽

H. Greifová ◽

A. Mackovich ◽

F. Hashim ◽

N. Lukáč

Keyword(s):

Oxidative Stress ◽

Oxidative Damage ◽

Structural Integrity ◽

Mitochondrial Activity ◽

Stress Markers ◽

Male Gametes ◽

Bovine Spermatozoa ◽

Oxidative Damage To Proteins ◽

Antioxidant Molecule

Evidence shows that oxidative stress associated with sperm cryopreservation may lead to a significant decrease of the structural integrity and functional activity of male gametes. Curcumin (CUR) has become a substance of scientific interest for its free radical-quenching abilities, which could enhance the post-thaw quality of male gametes. This study assessed the effects of CUR on the post-thaw vitality and selected oxidative stress markers of bovine spermatozoa. Thirty ejaculates collected from 10 breeding bulls were divided into two aliquots and cryopreserved in the absence (control) or presence of CUR (50 μmol/l). Immediately before use, the control or experimental straws were thawed at 37°C for 20 s. CUR administration led to a significantly higher preservation of spermatozoa motion (P < 0.001) as well as membrane (P < 0.05) and acrosomal (P < 0.01) integrity in comparison with the control. Moreover, spermatozoa exposed to CUR exhibited a significantly higher mitochondrial activity (P < 0.001). Significantly decreased amounts of reactive oxygen species (P < 0.01) and superoxide (P < 0.001) were detected following CUR supplementation. Finally, a significant decrease of oxidative damage to proteins (P < 0.01), lipids (P < 0.001), and DNA (P < 0.05) was recorded in samples to which CUR was administered in comparison to the control. In this study, CUR proved to act as an efficient antioxidant molecule offering protection to male gametes against oxidative damage during cryopreservation.

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Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates

Reproduction Fertility and Development ◽

10.1071/rd15261 ◽

2017 ◽

Vol 29 (6) ◽

pp. 1107 ◽

Cited By ~ 12

Author(s):

C. M. Kershaw ◽

G. Evans ◽

R. Rodney ◽

W. M. C. Maxwell

Keyword(s):

Sperm Motility ◽

Exposure Time ◽

Seminal Plasma ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Plasma Viscosity ◽

Dna Integrity ◽

Sperm Function ◽

Viscous Component ◽

Acrosome Integrity

In camelids, the development of assisted reproductive technologies is impaired by the viscous nature of the semen. The protease papain has shown promise in reducing viscosity, although its effect on sperm integrity is unknown. The present study determined the optimal papain concentration and exposure time to reduce seminal plasma viscosity and investigated the effect of papain and its inhibitor E-64 on sperm function and cryopreservation in alpacas. Papain (0.1 mg mL–1, 20 min, 37°C) eliminated alpaca semen viscosity while maintaining sperm motility, viability, acrosome integrity and DNA integrity. Furthermore E-64 (10 µM at 37°C for 5 min after 20 min papain) inhibited the papain without impairing sperm function. Cryopreserved, papain-treated alpaca spermatozoa exhibited higher total motility rates after chilling and 0 and 1 h after thawing compared with control (untreated) samples. Papain treatment, followed by inhibition of papain with E-64, is effective in reducing alpaca seminal plasma viscosity without impairing sperm integrity and improves post-thaw motility rates of cryopreserved alpaca spermatozoa. The use of the combination of papain and E-64 to eliminate the viscous component of camelid semen may aid the development of assisted reproductive technologies in camelids.

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Production and scavenging of reactive oxygen species both affect reproductive success in male and female Drosophila melanogaster

Biogerontology ◽

10.1007/s10522-021-09922-1 ◽

2021 ◽

Author(s):

Biz R. Turnell ◽

Luisa Kumpitsch ◽

Klaus Reinhardt

Keyword(s):

Reactive Oxygen Species ◽

Drosophila Melanogaster ◽

Reactive Oxygen ◽

Cellular Aging ◽

Ros Production ◽

Oxygen Species ◽

Wild Type ◽

Ros Scavenging ◽

Respiratory Pathway ◽

Male And Female

AbstractSperm aging is accelerated by the buildup of reactive oxygen species (ROS), which cause oxidative damage to various cellular components. Aging can be slowed by limiting the production of mitochondrial ROS and by increasing the production of antioxidants, both of which can be generated in the sperm cell itself or in the surrounding somatic tissues of the male and female reproductive tracts. However, few studies have compared the separate contributions of ROS production and ROS scavenging to sperm aging, or to cellular aging in general. We measured reproductive fitness in two lines of Drosophila melanogaster genetically engineered to (1) produce fewer ROS via expression of alternative oxidase (AOX), an alternative respiratory pathway; or (2) scavenge fewer ROS due to a loss-of-function mutation in the antioxidant gene dj-1β. Wild-type females mated to AOX males had increased fecundity and longer fertility durations, consistent with slower aging in AOX sperm. Contrary to expectations, fitness was not reduced in wild-type females mated to dj-1β males. Fecundity and fertility duration were increased in AOX and decreased in dj-1β females, indicating that female ROS levels may affect aging rates in stored sperm and/or eggs. Finally, we found evidence that accelerated aging in dj-1β sperm may have selected for more frequent mating. Our results help to clarify the relative roles of ROS production and ROS scavenging in the male and female reproductive systems.

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Somatic production of reactive oxygen species does not predict its production in sperm cells across Drosophila melanogaster lines

BMC Research Notes ◽

10.1186/s13104-021-05550-7 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Biz R. Turnell ◽

Luisa Kumpitsch ◽

Anne-Cécile Ribou ◽

Klaus Reinhardt

Keyword(s):

Reactive Oxygen Species ◽

Drosophila Melanogaster ◽

Reactive Oxygen ◽

Future Research ◽

Ros Production ◽

Oxygen Species ◽

Loss Of Function ◽

Wild Type ◽

Ros Scavenging ◽

Transport Chain

Abstract Objective Sperm ageing has major evolutionary implications but has received comparatively little attention. Ageing in sperm and other cells is driven largely by oxidative damage from reactive oxygen species (ROS) generated by the mitochondria. Rates of organismal ageing differ across species and are theorized to be linked to somatic ROS levels. However, it is unknown whether sperm ageing rates are correlated with organismal ageing rates. Here, we investigate this question by comparing sperm ROS production in four lines of Drosophila melanogaster that have previously been shown to differ in somatic mitochondrial ROS production, including two commonly used wild-type lines and two lines with genetic modifications standardly used in ageing research. Results Somatic ROS production was previously shown to be lower in wild-type Oregon-R than in wild-type Dahomey flies; decreased by the expression of alternative oxidase (AOX), a protein that shortens the electron transport chain; and increased by a loss-of-function mutation in dj-1β, a gene involved in ROS scavenging. Contrary to predictions, we found no differences among these four lines in the rate of sperm ROS production. We discuss the implications of our results, the limitations of our study, and possible directions for future research.

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Molecular Mechanisms Responsible for Increased Vulnerability of the Ageing Oocyte to Oxidative Damage

Oxidative Medicine and Cellular Longevity ◽

10.1155/2017/4015874 ◽

2017 ◽

Vol 2017 ◽

pp. 1-22 ◽

Cited By ~ 25

Author(s):

Bettina P. Mihalas ◽

Kate A. Redgrove ◽

Eileen A. McLaughlin ◽

Brett Nixon

Keyword(s):

Oxidative Damage ◽

Molecular Mechanisms ◽

Spindle Assembly Checkpoint ◽

Assisted Reproductive Technologies ◽

Reproductive Technologies ◽

Oocyte Quality ◽

Protein Repair ◽

Protective Mechanisms ◽

Pronounced Increase ◽

Age Related

In their midthirties, women experience a decline in fertility, coupled to a pronounced increase in the risk of aneuploidy, miscarriage, and birth defects. Although the aetiology of such pathologies are complex, a causative relationship between the age-related decline in oocyte quality and oxidative stress (OS) is now well established. What remains less certain are the molecular mechanisms governing the increased vulnerability of the aged oocyte to oxidative damage. In this review, we explore the reduced capacity of the ageing oocyte to mitigate macromolecular damage arising from oxidative insults and highlight the dramatic consequences for oocyte quality and female fertility. Indeed, while oocytes are typically endowed with a comprehensive suite of molecular mechanisms to moderate oxidative damage and thus ensure the fidelity of the germline, there is increasing recognition that the efficacy of such protective mechanisms undergoes an age-related decline. For instance, impaired reactive oxygen species metabolism, decreased DNA repair, reduced sensitivity of the spindle assembly checkpoint, and decreased capacity for protein repair and degradation collectively render the aged oocyte acutely vulnerable to OS and limits their capacity to recover from exposure to such insults. We also highlight the inadequacies of our current armoury of assisted reproductive technologies to combat age-related female infertility, emphasising the need for further research into mechanisms underpinning the functional deterioration of the ageing oocyte.

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Will the Bacteria Survive in the CeO2 Nanozyme-H2O2 System?

Molecules ◽

10.3390/molecules26123747 ◽

2021 ◽

Vol 26 (12) ◽

pp. 3747

Author(s):

Weisheng Zhu ◽

Luyao Wang ◽

Qisi Li ◽

Lizhi Jiao ◽

Xiaokan Yu ◽

...

Keyword(s):

Peroxidase Activity ◽

Biomedical Applications ◽

Antibacterial Effect ◽

Ros Production ◽

Ros Scavenging ◽

Ros Scavenger ◽

Oxide Nanoparticle ◽

H2o2 System ◽

Bacterial Disinfection

As one of the nanostructures with enzyme-like activity, nanozymes have recently attracted extensive attention for their biomedical applications, especially for bacterial disinfection treatment. Nanozymes with high peroxidase activity are considered to be excellent candidates for building bacterial disinfection systems (nanozyme-H2O2), in which the nanozyme will promote the generation of ROS to kill bacteria based on the decomposition of H2O2. According to this criterion, a cerium oxide nanoparticle (Nanoceria, CeO2, a classical nanozyme with high peroxidase activity)-based nanozyme-H2O2 system would be very efficient for bacterial disinfection. However, CeO2 is a nanozyme with multiple enzyme-like activities. In addition to high peroxidase activity, CeO2 nanozymes also possess high superoxide dismutase activity and antioxidant activity, which can act as a ROS scavenger. Considering the fact that CeO2 nanozymes have both the activity to promote ROS production and the opposite activity for ROS scavenging, it is worth exploring which activity will play the dominating role in the CeO2-H2O2 system, as well as whether it will protect bacteria or produce an antibacterial effect. In this work, we focused on this discussion to unveil the role of CeO2 in the CeO2-H2O2 system, so that it can provide valuable knowledge for the design of a nanozyme-H2O2-based antibacterial system.

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