| In order to study the effect of chlorogenic acid,luteolin and Polygonatum Polysaccharide on the cryopreservation of porcine semen,select the appropriate dosage and guide the production practice.In this study,chlorogenic acid,luteolin and Polygonatum Polysaccharide were used as protective additives in the diluent of boar semen cryopreservation.The effects of cryopreservation on sperm were determined by measuring the sperm motility,viability,plasma membrane,acrosome and DNA integrity,mitochondrial,superoxide dismutase,catalase and glutathione peroxidase activities.Chlorogenic acid,luteolin and Polygonatum Polysaccharide were added in the experiment,and the optimal dosage and compatibility were selected,which were applied to the experiment of boar semen freezing and artificial insemination.1.The results showed that the addition of chlorogenic acid had a certain improvement on the detection indexes of boar sperm after freezing thawing,among which the 50 ?g/ml test group had the best effect(P< 0.05).However,there was no strong correlation between the concentration of chlorogenic acid and sperm motility,acrosome,plasma membrane,DNA integrity and mitochondrial activity.The sperm activity,acrosome and membrane integrity rate of the 15 ?g/ml group were not significantly different from those in the control group(P> 0.05).The dosage of the addition was related to the enzyme activity of the antioxidant enzyme of the semen after freezing.The SOD,CAT and GSH-Px activities of the group were increased from 30 ?g/ml to 100 ?g/ml(P< 0.05).The results showed that the activity of CAT and GSH-Px in the 15 ?g/ml group was not significantly different(P> 0.05).The results showed that the addition of chlorogenic acid in the frozen diluent of boar semen had a certain effect on the activity rate,membrane,integrity and DNA integrity rate of frozen sperm,and the activity of mitochondria,SOD,CAT and GSH-Px,and the best dosage was 50 ?g/ml.2.Luteolin has a certain effect on the detection indexes of frozen thawed boar sperm.There is a weak positive correlation between the dosage and the motility of frozen sperm,the integrity rate of acrosome,plasma membrane and DNA,and the activity of mitochondria.When 80 ?g/ml was added,the plasma membrane integrity rate was significantly higher than that of the control group(P< 0.01),and the antioxidant enzyme activity of the experimental group was significantly higher than that of the control group(P< 0.05).When 100 ?g/ml sperm was added,the activities of SOD,CAT and GSH-Px decreased again,and there was no significant difference between the experimental group and the control group(P> 0.05).The results showed that luteolin could improve sperm motility,plasma membrane integrity,DNA integrity,mitochondrial,SOD,CAT and GSH-Px activities,and the optimal dosage was 80 ?g/ml.The addition of Polygonatum Polysaccharide could improve the viability of frozen thawed pig sperm,and the addition of 30 ? g/m L?80 ? There was significant difference between the treatment group and the control group(P < 0.05)? g/m L?50? g/m L?100 ? There was no significant difference between the two groups(P >0.05).The peak values of sperm motility,plasma membrane and DNA integrity all appeared at 30 ? The peak of acrosome integrity rate appeared at 80 g / ml ? G / ml,and 50 ? There was significant difference between the two groups(P < 0.05)3.The results showed that Polygonatum Polysaccharide could improve the viability of porcine sperm after freezing to a certain extent,and there was significant difference between the treatment groups with 30 ?g/ml and 80 ?g/ml and the control group(P< 0.05),while there was no significant difference between the treatment groups with 15 ?g/ml,50 ?g/ml and 100 ?g/ml and the control group(P> 0.05).The peak values of sperm motility,plasma membrane and DNA integrity all appeared in the 30 ?g/ml group,and the peak value of acrosome integrity appeared in the 80?g/ml group,which was significantly different from the 50 ?g/ml group(P< 0.05),and the difference between the experimental group and the control group was extremely significant(P< 0.01).There was no significant correlation between the dosage and sperm motility and functional integrity,and there was no significant difference between 50 ?g/ml and 100 ?g/ml groups(P> 0.05).The results showed that Polygonatum Polysaccharide could improve the antioxidant enzyme activity of frozen sperm,and the 30 ?g/ml treatment group was significantly different from the control group(P< 0.05).The peak of SOD activity of sperm in 80 ?g/ml treatment group was significantly higher than that in the control group(P< 0.05);the peak of GSH-Px activity of sperm in 30 ?g/ml treatment group was significantly higher than that in the control group(P< 0.01);but the activities of SOD,CAT and GSH-Px in15 ?g/ml treatment group were not significantly different from that in the control group(P> 0.05).The results showed that the proper amount of Polygonatum Polysaccharide added to the frozen diluent of boar semen had a certain effect on sperm motility,plasma membrane,acrosome and DNA integrity,mitochondrial,SOD,CAT and GSH-Px activities,and the optimal amount was 30 ?g/ml.4.There were significant differences between the different compatibility groups and the control group(P< 0.05),which could improve the viability,acrosome,plasma membrane and DNA integrity of porcine sperm after freezing.Among them,the compatibility group VI(35 ?g/ml chlorogenic acid plus 40 ?g/ml luteolin plus 15?g/ml Polygonatum Polysaccharide)had the best effect,with the highest sperm motility,plasma membrane,acrosome and DNA integrity rates of 48.9%,50.81%,50.09%,49.36% and 48.13%,respectively.There were significant differences in sperm motility,acrosome and plasma membrane integrity among the compatibility groups,but there was no significant difference in sperm DNA integrity among the compatibility groups(P> 0.05).There was significant difference between the different compatibility groups and the control group(P< 0.05),which indicated that the different compatibility groups could improve the antioxidant enzyme activity of frozen pig sperm,and there was a certain synergistic effect.Among them,the compatibility group VI had the best effect,and the highest SOD,CAT and GSH-Px were 69.79 U/ml,3.98 U/ml and 221.37 U/I,respectively.The results showed that different compatibility of chlorogenic acid,luteolin and Polygonatum Polysaccharide in pig semen cryodiluent had certain effects on sperm motility,plasma membrane,acrosome integrity,DNA integrity,mitochondrial,SOD,CAT and GSH-Px activity.The best compatibility was 35 ?g/ml chlorogenic acid,40 ?g/ml luteolin and 15?g/ml Polygonatum Polysaccharide.5.Compared with artificial insemination with fresh boar semen,frozen boar semen had lower rates of no return to estrus,conception,delivery and litter size.But in the frozen semen test group,the results of each treatment group were significantly better than that of the control group.After adding 50 ?g/ml chlorogenic acid,80?g/ml luteolin and 30 ?g/ml Polygonatum Polysaccharide to the frozen diluent of boar semen,and adding 35 ?g/ml chlorogenic acid,40 ?g/ml luteolin and 15 ?g/ml Polygonatum Polysaccharide together,litter size was significantly increased(P<0.05).The results showed that compared with the results of artificial insemination of fresh boar semen,the effect of artificial insemination of frozen semen was still not ideal,but in each frozen experimental group,50 ?g/ml chlorogenic acid,80 ?g/ml luteolin,30 ?g/ml Polygonatum Polysaccharide and 35 ?g/ml chlorogenic acid,40?g/ml luteolin,15 ?g/ml Polygonatum Polysaccharide test group has a certain effect on improving the monitoring index of artificial insemination.The litter size of compatibility group was significantly higher than that of other frozen groups(P<0.05).In conclusion,adding Chlorogenic acid,Luteolin and Polygonatum Polysaccharide in the cryodiluent has a certain effect on the cryoprotection and artificial insemination of boar semen,and the best effect is the compatibility group. |
PDF Full Text Request