The Effects Of Three Antioxidants Of NAC、SLS And SDS On Frozen-thawed Sheep Sperm

The large amount of active oxygen produced during the cryopreservation of semen will damage the anti-oxidant system of the sperm,thereby inducing sperm apoptosis and making the sperm lose the ability to fertilize.Therefore,the method of adding antioxidants to semen cryopreservation diluent is usually used to improve the effect of semen cryopreservation.N-Acetyl Cysteine(NAC)and SodiumLauryl Sulfonate(SLS)have been proven to play a protective role in the cryopreservation of pig semen.SodiumLauryl Sulfate(SDS)can improve canine sperm cryopreservation After the survival rate.However,at present,the role of these three antioxidants on goat semen cryopreservation is still unclear.In this experiment,the Saanen dairy goat was used as the experimental object to analyze the effects of NAC,SLS and SDS on the freezing and thawing of Saanen dairy goat semen before and after freezing,so that these three types of antioxidants can be used in the Sanen dairy goat semen cryopreservation technology.The application provides theoretical basis and technical support.The main results of this experiment are as follows:1.Adding different concentrations of NAC(0.2,0.4,0.6,0.8,1.0 mg/mL)to the frozen storage diluent of Sanen dairy goat semen has a positive effect on the quality of frozen-thawed semen.When the concentration of NAC in the diluent is 0.6 mg/mL,the sperm viability and plasma membrane intact rate after freezing-thawing are 43.19% and48.52%,respectively;the total anti-oxidant capacity(T-AOC)of sperm reaches 9.93IU/mL,Superoxide dismutase(SOD)is 220.3IU/mL,catalase(CAT)is 8.53IU/mL,acid phosphatase(ACP)is 33.14IU/L,glutathione peroxidase(GSH-Px)reached 135.4IU/mL,hyaluronidase(HA)was 355.17IU/L,sperm acrosin(ACE)was 5.074IU/L,which were significantly higher than those of the control group(P<0.05).At the same time,the level of metabolite reactive oxygen species(ROS)was 2968 and malondialdehyde(MDA)was5.241 mm/mL,both of which were significantly lower than the control group(P<0.05).The mitochondrial membrane potential was 2.993 Δ Ψm,which was significantly higher than that of the control group(P<0.05).2.Adding different concentrations of SLS(0.05,0.1,0.15,0.2,0.25 mg/mL)to the frozen storage diluent of Sanen dairy goat semen has a positive effect on the quality of semen after freezing and thawing.When the amount of SLS added is 0.15 mg/mL,the sperm viability and plasma membrane intact rate after freezing and thawing are 45.03% and46.72% respectively;the total antioxidant capacity(T-AOC)of sperm reaches 8.90IU/mL,superoxide Dismutase(SOD)is 201.3 IU/mL,acid phosphatase(ACP)is 34.04IU/L,catalase(CAT)is 8.23IU/mL,and glutathione peroxidase(GSH-Px)is 162.2IU/mL,hyaluronidase(HA)421.15 IU/L,sperm acrosin(ACE)was 5.476 IU/L,which were significantly higher than those of the control group(P<0.05).At the same time,the level of metabolite reactive oxygen species(ROS)was 3739 and malondialdehyde(MDA)was5.345 mm/mL,both of which were significantly lower than the control group(P<0.05).The mitochondrial membrane potential was 2.983 Δ Ψm,which was significantly higher than that of the control group(P<0.05).3.The addition of different concentrations of SDS(0.05,0.1,0.15,0.2,0.25 mg/mL)to the frozen storage diluent of Sanen dairy goat semen has a positive effect on the quality of semen after freezing and thawing.After adding 0.15 mg/mL SDS,the sperm viability and plasma membrane intact rate after freezing and thawing were 46.01% and 42.28%,respectively;the total anti-oxidant capacity(T-AOC)of sperm reached 7.30IU/mL,superoxide dismutation Enzyme(SOD)is 202.0 IU/mL,catalase(CAT)is 8.46 IU/mL,acid phosphatase(ACP)is 34.47 IU/L,and glutathione peroxidase(GSH-Px)is 175.92 IU/mL,hyaluronidase(HA)452.72 IU/L,sperm acrosin(ACE)was 5.793 IU/L,which were significantly higher than those of the control group(P<0.05).At the same time,the level of metabolite reactive oxygen species(ROS)was 3320 and malondialdehyde(MDA)was5.476 mm/mL,both of which were significantly lower than the control group(P<0.05).The mitochondrial membrane potential was 2.750 Δ Ψm,which was significantly higher than that of the control group(P<0.05).Studies have shown that adding NAC,SLS and SDS to sheep semen freezing diluent can reduce the oxidative damage of sperm during freezing and improve the quality of frozen semen.At the same time,this study also optimized the optimal concentration.N-acetylcysteine is 0.6 mg/mL,sodium lauryl sulfonate is 0.15mg/mL,and sodium lauryl sulfate is 0.15 mg/mL.