Effect Of The Addition Of Melatonin,Readuced Glutathione And Vitamine C To The Boar Semen Extenders

The present research studied the peroxidation damage of boar semen and the effect of addition of antioxidants on the semen quality in cryopreservation and intend to reveal the mechanism of peroxidation damage and screening better antioxitants added to the extender thus improve the quality of boar semen after freezen-thawed.The change of ROS in cryopreservation was detected by primary fluorescence detection kit. After equilibrium,the content of ROS slightly increased;after freezen-thawed,the content of ROS significantly increased(P＜0.01).Detect the quality of fresh,after equilibrium,freezen-thawed semen respectively,we found that the quality of semen decreased significantly(P＜0.01).In addition,the negative correlation between the content of ROS and quality of boar semen was significantly through the use of SPSS 13.0,the correlation coefficient of viability,motility,acrosomal integrity,and mitochondrial activity were -0.879,-0.958,-0.819,-0.960 respectively.The centrifugation tested were 2400×g for 3min and 800×g for 10min.,and we recommend the use of short-term centrifugation with a relatively high g-force(2400 3 g for 3 minutes) in boar sperm cryopreservation protocol,it can improve the motility,viability,and mitochondrial activity significantly(P＜0.05).The addition of melatonin(0.125mg/ml,0.25 mg/ml,0.5 mg/ml) to the boar semen extender was the first time,when added 0.25 mg/ml melatonin can improve the motility,mitochondrial activity,and acrosomal integrity significantly(P＜0.05).Added the readuced glutathione(lmmol/L,5 mmol/L,10 mmol/L)to boar semen extenders,5 mmol/L can improve motility,viability and mitochondrial activity significantly (P＜0.01).both the 1 mmol/L and 5 mmol/L can improve the acrosomal integrity significantly(P＜0.05),but there was no significant difference between the two concentration.Use the orthogonal design first time to study the associated addition of melatonin,readuced glutathione and vitamine C.Through analysis,wo found that the interaction between melatonin and readuced glutathione was notablely(P＜0.05),and the interaction of other combination was not noteblely(P＜0.05).Morever,we should avoid the presense of Fe²⁺and Cu⁺ when add vitamine C to the extenders,in order to avoid catalysis the oxidation of vitamine C to produce H₂O₂ and damage plasmalemma.Study the interaction between melatonin and readuced glutathione fuether,we found that added melatonin 0.125mg/ml,readuced glutathione lmmol/L can improve the motility,viability mitochondrial activity,and acrosomal integrity notablely(P＜0.05).The effect of associated addition of melatonin,readuced glutathione was better than added independently.After all,we recommend use centrifugation:2400×g for 3min,and add melatonin 0.125mg/ml,readuced glutathione 1mmol/L can obtain a good result.