Effects of Mito-TEMPO on cryopreservation of porcine spermatozoa
[Objective]The study aimed to select the optimal concentration of Mito-TEMPO.Meanwhile,the effect of Mito-TEMPO on the quality,antioxidation capacity,and mitochondrial function of frozen-thawed porcine spermatozoa were explored to provide a reference basis for the improvement of porcine spermatozoa cryopreservation.[Method]0.5,5.0,50.0 and 500.0 μmol/L Mito-TEMPO were added to the frozen solution Ⅰ in the experimental group.The concentration of the Mito-TEMPO suitable for the cry opreservation of porcine spermatozoa was screened by evaluating the kinetic parameters and viability of spermatozoa in each group.The plasma membrane intactness,acrosome in-tactness,DN A fragmentation index(DFI),reactive oxygen species(ROS)level,total antioxidant capacity(T-AOC),malondialdehyde(MDA)content,catalase(CAT)and total superoxide dismutase(T-SOD)activities,as well as ATP and mitochondrial respiratory chain complex Ⅰ-Ⅴ content of freezing-thawing porcine spermatozoa were detected.[Result]Compared to the fresh group(F group),the kinetic parameters and viability of spermatozoa were significantly reduced after freezing-thawing(P＜0.05,the same as below).While the addition of Mito-TEMPO improved the kinetic parameters and viability of frozen-thawed spermatozoa to various degrees,and the effect of 50.0 μmol/L Mito-TEMPO was the optimal.Compared to the F group,the plasma membrane and acrosome integrity,T-AOC,ATP and mitochondrial respirato-ry chain complex Ⅰ-Ⅴ contents of frozen-thawed spermatozoa were significantly reduced,as well as CAT and T-SOD activities.DFI,ROS levels and MDA contents were significantly increased.Compared to the frozen control group(C group),the plasma membrane integrity,ac-rosome integrity,T-AOC,as well as ATP and mitochondrial respiratory chain complex Ⅰ,Ⅱ and Ⅴ contents of spermatozoa were significant-ly increased in the 50.0 μmol/L Mito-TEMPO group(M group).Moreover,the DFI,ROS levels and MDA contents were significantly re-duced,and the CAT and T-SOD activities were significantly enhanced by the 50.0 μmol/L Mito-TEMPO.[Conclusion]Mito-TEMPO has a cryoprotective effect on porcine spermatozoa,and the optimal concentration is 50.0 μmol/L.In addition,50.0 μmol/L Mito-TEMPO in-creases the quality of spermatozoa by improving the antioxidation capacity and mitochondrial function of porcine spermatozoa after freezing-thawing.
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