Effect Of Proline On Boar Semen Quality During Normal Temperature Preservation

Mammalian spermatozoa are extremely susceptible to high doses of reactive oxygen species?ROS?.The aim of the present study was to investigate the potential role of proline in protecting boar spermatozoa against ROS stress during ambient temperature storage,and if so,how proline protects spermatozoa from the damage during the preservation.Fresh semen was diluted with Modena extender supplemented with different concentrations of proline.The motility,intact acrosomes,membrane integrity,mitochondrial potentials were examined.Furthermore,the level of reactive oxygen species?ROS?,lipid peroxidation?LPO?,glutathione content?GSH?,the activity of catalase?CAT?and superoxide dismutase?SOD?were analyzed.Besides,we also evaluated the content of ATP.The results are shown as follows:1.The values of total motility?%?,progressive motility?%?,straight-line velocity(VSL,?m s^-1),curvilinear velocity(VCL,?m s^-1),average velocity(VAP,?m s^-1),straightness?STR,ratio of VSL/VAP,%?,wobble of the curvilinear trajectory?WOB,ratio of VAP/VCL,%?and beat cross-frequency?BCF,Hz?,intact acrosomes,membrane integrity,mitochondrial potentials of the boar spermatozoa in the treatment of 75 mmol/L proline were significantly higher than those of the control?P<0.05?.2.The levels of ROS and lipid peroxidation were the lowest in 75 mmol/L proline treatment group.By comparing with the control group,the levels of glutathione content?GSH?,the activity of catalase?CAT?and superoxide dismutase?SOD?were significantly higher?P<0.05?.The results showed that proline could protect spermatozoa by removing spermatozoa ROS,reduced the level of lipid peroxidation,improved the capacity of CAT and SOD and alleviate the oxidation damage during the normal temperature preservation.The results of western blot showed that proline could decrease the apoptosis of treatment sperm.Meanwhile the presence of proline dehydrogenase in spermatozoa indicates that proline may be involved in the metabolism of sperm cells and provides ATP for the preservation process of porcine semen at normal temperature.3.In semen by adding hydrogen peroxide 37?were incubated with 2 h,total sperm motility,linear motion ability CASA detection the boar semen quality parameters and plasma membrane and acrosome integrity,mitochondrial activity decreased significantly?P<0.05?,the level of lipid peroxidation and ROS increased significantly?P<0.05?.Boar semen quality parameters CASA showed significantly increased?P<0.05?,plasma membrane and acrosome integrity,mitochondrial activity was significantly increased?P<0.05?,the level of lipid peroxidation and ROS decreased significantly?P<0.05?with the treatment of 75 mmol/L proline,further it shows that proline through their antioxidant ability to protect the boar sperm from.In summary,this study shows that proline can be through its antioxidant capacity of boar semen preservation play a protective role at a concentration of 75 mmol/L;at the same time,proline may also be involved in sperm Cell Metabolismolism,provided ATP for cell survival,improving the quality of semen preservation.