Study Of ZnO NPs Improved The Quality Of Liquid-storage Boar Sperm

The liquid storage of boar sperm is an important step in the implementation of artificial insemination（AI）technology,which has the ability to increase the lifespan of sperm in vitro and maintain sperm fertility.The liquid storage of boar sperm is important for increasing boar utilization and improving genetic breeding of boar.Sperm quality can be negatively impacted by oxidative stress,DNA strand breakage,and lipid peroxidation because of the accumulation of ROS during storage.Determining the extender suitable for the liquid storage of boar sperm is therefore crucial for the advancement of AI.Furthermore,adding antioxidants can limit ROS generation and increase sperm quality.Zinc oxide nanoparticles（Zn O NPs）,a new source of zinc,exhibit potent antioxidant properties.It is yet unknown what influence Zn O NPs have on the liquid storage of boar sperm.In this experiment,Zn O NPs were added to the extender for liquid（4°C,17°C）storage.Using the computer-aided sperm analysis（CASA）system analysis,western blotting,and other experimental techniques,the motility,membrane integrity,and protein phosphorylation levels,and other parameters of sperm were detected.This research investigated the effect of Zn O NPs on the liquid storage of boar sperm.The following were the main outcomes.1.To determine the extender suitable for liquid storage of boar sperm,different concentrations（0.1μg/m L,1μg/m L,10μg/m L）of Zn O NPs were added to the basic extender for liquid storage,and boar sperm motility,viability,membrane integrity,acrosome integrity,and abnormality rate were measured.The results showed that the 0.1μg/m L Zn O NPs treatment group’s sperm motility,viability,membrane integrity,acrosome integrity,and abnormality rate were not significantly different from the control group.Boar sperm motility（79.27 at 4℃and 95.67%at 17℃）,viability（69.24%at 4℃and 78.95%at 17℃）,membrane integrity（79.27%at 4℃and 86.67%at 17℃）,and acrosome integrity（79.67%at 4℃and93.27%at 17℃）were significantly greater in the 1μg/m L Zn O NPs treatment group than in the control group,but there was no significant difference in abnormality rate.Boar sperm motility,viability,membrane integrity,and acrosome integrity did not vary significantly from the control group in the 10μg/m L Zn O NPs treatment group,but the abnormality rate of boar sperm stored at 17°C（12.56%）was significantly higher than in the control group.According to the above findings,adding 1μg/m L Zn O NPs could improve the quality of liquid storage boar sperm,whereas adding 10μg/m L Zn O NPs was detrimental to the morphology of liquid storage boar sperm.As a result,in following experiments,1μg/m L Zn O NPs were used for treatment.2.To investigate the impact of Zn O NPs on the antioxidant level of liquid storage boar sperm,the experiment added 1μg/m L Zn O NPs to the extender for liquid storage and measured MMP,ROS,malondialdehyde（MDA）,and other antioxidant levels in sperm.The results showed that,when compared to the control group,ROS and MDA levels were significantly lower in the Zn O NPs treatment group,while mitochondrial membrane potential（MMP）,the total antioxidant capacity（T-AOC）,the activities of superoxide dismutase（SOD）,glutathione peroxidase（GSH-Px）were significantly higher.The findings suggested that Zn O NPs can act as antioxidants during liquid storage.3.Protein Serine/threonine phosphorylation（P-PKAs）and tyrosine phosphorylation（PTP）levels of liquid storage of boar sperm were detected in the research to figure out the mechanism of Zn O NPs improving the quality of liquid storage of boar sperm.The results revealed that Zn O NPs could significantly improve the level of P-PKAs in boar sperm liquid storage but had no effect on the level of PTP.Hydrogen peroxide（H₂O₂）was used to build an oxidative stress model in order to further investigate the process of Zn O NPs in alleviating oxidative stress of liquid storage of boar sperm.The findings indicate that Zn O NPs could significantly improve the level of P-PKAs in boar sperm induced by ROS,so as to reduce oxidative stress in sperm.In conclusion,Zn O NPs can protect boar sperm by lowering oxidative stress in boar sperm during liquid storage,mitigate sperm quality,improve the duration of liquid storage,and increase storage effectiveness in vitro.1μg/m L is the appropriate addition concentration.