Effect Of Antioxidants GLP And Mito Q On The Cryopreservation Of Goat Semen

The effective combination of cryopreservation of semen and artificial insemination has a positive effect on the protection of germplasm resources,production and breeding,etc.However,in the process of cryopreservation of semen,sperm cells are extremely vulnerable to different degrees of physical damage,chemical damage and oxidative stress.Excitation damage.Among them,oxidative stress damage is the main factor that damages sperm in the process of cooling and preservation,which will have a series of adverse effects on the structure and morphology of sperm,such as: destroying sperm structure,reducing the sperm plasma membrane,acrosome,DNA and mitochondria after freezing-thawing The integrity of the sperm cells causes the sperm cells to lose their normal physiological functions and reduces the quality of semen after freezing and thawing.A large number of in vitro studies have found that during the cryopreservation of semen,the addition of exogenous antioxidants is a key factor in reducing oxidative stress and reducing ROS levels.Ganoderma lucidum polysaccharides(GLP)is a carbohydrate antioxidant.In the process of semen cryopreservation,it can not only act as an antioxidant to protect sperm from oxidative stress,but also provide nutrients for semen cryopreservation;Mitoxene Quinone mesylate(Mitoquinone,Mito Q)is a mitochondrial targeted antioxidant.Studies have found that Mito Q plays a protective role in the cryopreservation of semen of fish,humans and sheep.This experiment took Saanen dairy goats as the research object,and explored the effect of adding two antioxidants(GLP and Mito Q)to the semen cryo-diluent on the cryopreservation of dairy goat semen.By testing the sperm motility,plasma membrane integrity,antioxidant capacity and acrosin activity after thawing,the optimal preservation concentration was screened.The test results are as follows:1.Add different concentrations of GLP(0.2,0.4,0.6,0.8,1.0 mg/m L)to the cryopreservation diluent.After freezing and thawing,the goat sperm quality test results found that the concentration of 0.8 mg/ml was added to the diluent.When m L,compared with the control group,sperm viability,plasma membrane integrity rate,and mitochondrial activity were significantly improved(P<0.05),sperm viability and plasma membrane integrity rate were 42.03 ± 0.71% and 50.51 ± 0.35%,respectively;When the added concentration was 1.0 mg/m L,the sperm viability after thawing was not significantly different from that of the control group(P> 0.05).In the test results of semen antioxidant capacity,it was found that the preservation effect was best when 0.8 mg/m L was added to the diluent.After freezing and thawing,the sperm T-AOC,SOD,CAT,GSH-PX were 8.17±0.18 U/m L,202.97±3.30 U/m L,3.23±0.16 U/m L,142.14±2.76 U/m L,ROS,MDA,respectively They were 3353 ± 68 and 4.48 ± 0.30 nmol/m L.According to the acrosin activity detection and analysis,it was found that when 0.8 mg/m L GLP was added,the preservation effect was the best(P<0.05).The ACP,HYD,and ACE were 32.78 ±,respectively.0.67 U/m L,386.92±5.07 U/L,6.25±0.04 U/L.The preservation effect of adding 0.6 mg/m L GLP was the second.When the concentration was 1.0 mg/m L,the acrosin enzyme activity decreased slightly,but they were all higher than the control group.2.Add different concentrations of Mito Q(50,100,150,200,250 nmol/L)to the frozen diluent.After freezing and thawing,the goat semen quality test results found that: the appropriate concentration of Mito Q can freeze goat semen Preservation has a protective effect.When the added concentration is 150 nmol/L,the sperm viability,plasma membrane intact rate,and mitochondrial activity after thawing are significantly higher than those of the control group(P<0.05).When the added concentration is greater than 200 nmol /L,it has no protective effect on semen cryopreservation.After thawing,sperm viability and plasma membrane integrity rate show a downward trend;in the test results of semen antioxidant capacity,it is found that 50 nmol/L-150 nmol/ is added to the cryopreservation diluent.When the concentration of L Mito Q increases,the preservation effect of semen is better.When the concentration is 150 nmol/L,the preservation effect is the best.The T-AOC,SOD,CAT,and GSH-PX are 7.87 ±0.20 U/L,respectively.m L,204.17 ±0.84 U/m L,8.99 ±0.21U/m L,142.27 ± 4.59 U/m L,ROS and MDA were 3149 ± 47.03 and 5.06 ± 0.14nmol/m L,respectively,which were significantly different from the control group(P <0.05);In the detection and analysis of semen acrosin activity,it was found that when Mito Q was added at a concentration of 150 nmol/L,there was a significant difference compared with the control group(P<0.05),and the ACP,HYD,and ACE were 30.51±0.44 U/ m L,334.42±3.60 U/L,6.57±0.08 U/L.However,as the concentration increased,the sperm acrosin activity showed a downward trend after thawing.When the added concentration was 250nmol/L,compared with the control group,the difference was not significant(P> 0.05).The results of this experiment show that when the concentration of 0.8 mg/m L GLP or the concentration of 150 nmol/L Mito Q is added to the frozen diluent of cashmere goat semen,the cryopreservation effect is the best,and the sperm viability and plasma membrane integrity rate after thawing are the best.,Antioxidant level and acrosin activity were significantly improved,and ROS and MDA in sperm were significantly reduced(P <0.05).