Mito-TEMPO 对猪精子冷冻保存效果的研究.

[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 cryopreservation of porcine spermatozoa was screened by evaluating the kinetic parameters and viability of spermatozoa in each group. The plasma membrane intactness, acrosome intactness, DNA 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 IV content of freezing-thawing porcine spermatozoa were detected. [Result] Compared to the fresh group (Fgroup), 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 respiratory chain complex IV 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 (Cgroup), the plasma membrane integrity, асrosome integrity, T-AOC, as well as ATP and mitochondrial respiratory chain complex I, II and V contents of spermatozoa were significantly increased in the 50.0 µmol/L Mito-TEMPO group (M group). Moreover, the DFI, ROS levels and MDA contents were significantly reduced, 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 increases the quality of spermatozoa by improving the antioxidation capacity and mitochondrial function of porcine spermatozoa after freezing-thawing. [ABSTRACT FROM AUTHOR]