The Effect Of Brassinolide,Diosmetin,and Ursolic Acid On Cryopreservation Of Boar Spermatozoa

Liquid semen preservation and frozen semen preservation are conventional techniques for gamete preservation of mammalian and domestic animals.It is well known that the decrease in temperature will reduce or stop the metabolic activity of cells,and the metabolic activity of cells will resume after warming.Liquid preservation can keep semen alive for a short time(from 24 hours to several days depending on the species),while cryopreservation is used for long-term preservation of semen and oocytes.Boar spermatozoa are sensitive to oxidative stress,so it is necessary to find suitable antioxidants to improve the quality of cryopreservation.Brassinolide,diosmetin,and ursolic acid have all been shown to be highly effective antioxidants and have never been used before to protect sperm from freezing damage.The purpose of this study was to investigate the effects of brassinolide,diosmetin,ursolic acid on sperm motility,viability,plasma membrane integrity,acrosome integrity,and mitochondrial membrane integrity in frozen boar semen.In this study,semen of five Landrace boars was used once a week.The results are as follows:1.Adding different concentrations of brassinolide to the frozen semen diluent can significantly improve sperm motility,viability,mitochondrial membrane integrity,plasma membrane integrity,and acrosome integrity.When the concentration of brassinolide was 1.8m M,the motility,activity,plasma membrane integrity,and acrosome integrity were the highest.However,when 1.6 mm brassinolide was added,the mitochondrial integrity was the highest.2.Adding different concentrations of diosmetin to frozen semen diluent can significantly improve sperm motility,viability,mitochondrial membrane integrity,plasma membrane integrity,and acrosome integrity.When the concentration of diosmetin was 1.2 m M,the sperm motility,viability,mitochondrial membrane integrity,plasma membrane integrity,and acrosome integrity of boar spermatozoa were the highest.3.Adding different concentrations of ursolic acid to the frozen semen diluent can significantly improve the sperm motility,viability,mitochondrial membrane integrity,plasma membrane integrity,acrosome integrity,and frozen-thawed semen integrity.When the concentration of ursolic acid was 1.6m M,the sperm viability,mitochondrial membrane integrity,plasma membrane integrity,and acrosome integrity were the highest.To sum up,adding antioxidants such as brassinolide,diosmetin,ursolic acid in the diluent can significantly improve sperm motility and viability,plasma membrane integrity,mitochondrial integrity,and acrosome integrity,and three substances have protective effects on frozen-thawed boar spermatozoa.