Study Of The Effect Of Puerarin On Pig Semen Preservation At Room Temperature

Artificial insemination technology has been widely used in the pig breeding industry.At present,the pig artificial insemination technology is mainly used with room temperature semen,which is stored under room temperature conditions,and is prone to oxidative stress,leading to the generation of a large amount of reactive oxygen species(ROS),which damages the membrane structure and function of sperm,and reduces the vitality and survival rate of sperm.Therefore,the study of semen preservation at normal temperature of pigs has a high practical value for the development of artificial insemination technology and even the production of pig industry.Studies have shown that the addition of natural protective agents can improve the preservation effect of pig semen at normal temperature.As a natural isoflavone antioxidant in nature,puerarin has the effect of improving biological immunity and protecting cardiomyocytes.At present,its effect in the preservation of pig semen has not been reported.Based on this,this study added different concentrations of puerarin to pig semen,detected sperm motility and motility parameters in the test group and blank group at the same time,selected the optimal concentration of puerarin,and further explored the effect of puerarin in the antioxidant capacity and energy conversion of pig semen.The main results are as follows:The concentrations of puerarin(0,10,25,50,70,100 μ mol/L),and sperm motility and sperm motility were determined by the computer(CASA)sperm analysis system showed that the mean motility was 81.6%,58.0%,and seventh day of the test group was 81.1%,70.6%,60 and 60.0%,respectively.Among them,the best sperm motility was in the 50 μmol/L puerarin supplementation group,with an average sperm motility of 83.8%,71.3%,and 64.3%.The mean sperm plasma membrane integrity and the control group were 70.4%,77.2%,73.8%,70.5%,75.4%,79.7% on the control group,and were 73.4%,61.2%,68.4%,71.8%,70.4% and 76%.Therefore,the pig sperm motility and sperm integrity rate in the 50 μ mol/L puerarin supplemented group were significantly higher than those in the control group(P <0.05).Aeffect of puerarin on the antioxidant capacity of pig semen: on the seventh day of semen storage at room temperature,The total antioxidant capacity(T-AOC)in the control semen was 1.95 n mol/L,The total antioxidant capacity(T-AOC)of 50 μ mol/L puerarin supplementation was 4.29 n mol/L,Significantly higher than that in the control group(P<0.05);Control reactive oxygen species(ROS)and malondialdehyde(MDA)levels were1.9106(AU)and 7.09 nmol / ml,respectively,And the 50 μ mol/L puerarin group was1.6106(AU)and 7.33 nmol / ml,respectively.Among them,ROS and MDA have the same trend in semen change,and the content of ROS and MDA shows an increasing trend,which is quite opposite to the change of T-AOC in semen.Seventh day,the catalase(CAT),superoxide dismutase(SOD),and glutathione peroxidase(GSH-PX)activities were 2.59(U / mg Hb),81.4(U / ml),3.40(μ mol/L),and 50 μ mol/L puerarin was 18.18(U / mg Hb),123.3(U / ml)and 12.39(μ mol/L),respectively,which was significantly higher than the control enzyme(P <0.05).In determining the effect of puerarin on sperm motility performance: in the sperm motility parameters(u m/sec)in the control group,the mean linear movement(VSL)was57.4,37.5,19.2,the mean path velocity(VAP)was 83.2,55.2,41.4,and the mean curve velocity(VCL)was 162.8,92.9 and 58.8,respectively.And the puerarin group of 50 μmol/L,VSL: 68.0,46.7,27.1,VAP: 95.1,67.7,53.9,and VCL: 173.4,114.5,70.4,respectively.By measuring the exercise parameters of VSL,VAP and VCL,the 50 μmol/L of puerarin swine semen group was significantly higher than that of the control group(P <0.05).Effect of puerarin on energy metabolism of pig semen: The control adenosine triphosphate(ATP)and lactate dehydrogenase(LDH)were 1772 nmol / 108 sperm and1.76(U / L),respectively,while the 50 μ mol/L puerarin addition group was 3219 nmol /108 sprint and 3.96(U / L),respectively.Therefore,the pig semen ATP level and LDH activity in the 50 μ mol/L puerarin group were significantly higher than those in the control group(P <0.05).