College of Life Science, Inner Mongolia University, 235 West Univ. Road, Hohhot, 010021, Inner Mongolia, China; Key Laboratory of Medical Cell Biology, Clinical Medicine Research Center, Affiliated Hospital of Inner Mongolia Medical University, 1 Tongdao North Street, Hohhot, 010050, Inner Mongolia, China.
College of Life Science, Inner Mongolia University, 235 West Univ. Road, Hohhot, 010021, Inner Mongolia, China.
Cryobiology. 2020 Dec;97:153-158. doi: 10.1016/j.cryobiol.2020.08.007. Epub 2020 Aug 26.
This present study investigated the effect of bioactive peptide (BAPT) (BAPT) on the quality of ram semen during cryopreservation. Ram ejaculates were extended with Tris buffer supplemented with no antioxidants (as control group), 20 μg/mL BAPT (as BAPT20 group), 40 μg/mL BAPT (as BAPT40 group) and 60 μg/mL BAPT (as BAPT60 group). After cryopreservation, sperm quality including motility, vitality, the percentage of hypoosmotic swelling test (HOST)-positive spermatozoa and the percentage of intact acrosomes was assessed. Furthermore, the malondialdehyde (MDA) in seminal plasma and spermatozoa were analyzed, followed by the measurement of superoxide dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GSH-Px) levels in seminal plasma. After in vitro fertilization, the embryonic cleavage rates and development rates of different groups were analyzed to compare the developmental abilities of spermatozoa. The results showed that the post-thaw sperm motility was significantly higher in the BAPT60 group compared to those in the BAPT20, BAPT40 and control groups (P < 0.05). The percentage of live sperms significantly increased from 48.12 ± 2.35% for the BAPT20 group, 55.43 ± 2.16% for the BAPT40 group to 57.53 ± 3.15% for the BAPT60 group. The percentage of HOST-positive spermatozoa was significantly higher in the BAPT60 group than those in BAPT20, BAPT40 and control groups (P < 0.05). The MDA levels in seminal plasma and spermatozoa were significantly reduced with BAPT supplement (P < 0.05). Additionally, the SOD, CAT and GSH-Px levels in the BAPT experimental groups were significantly higher than those of the control group, which further indicated that BAPT significantly inhibit the reactive oxygen species (ROS) production during the cryopreservation of ram semen. Furthermore, the embryonic cleavage rates and development rates of the BAPT40 and BAPT60 groups were significantly increased in comparison with the BAPT20 and control groups (P < 0.05). In conclusion, BAPT improved the ram sperm quality via inhibiting the ROS production during cryopreservation, and could be applied as a promising supplement for ram semen cryopreservation.
本研究旨在探讨生物活性肽（BAPT）对绵羊精液冷冻保存质量的影响。将绵羊精液用 Tris 缓冲液（不添加抗氧化剂，作为对照组）、20μg/mL BAPT（BAPT20 组）、40μg/mL BAPT（BAPT40 组）和 60μg/mL BAPT（BAPT60 组）进行延伸。冷冻保存后，评估精子质量，包括活力、活力、低渗肿胀试验（HOST）阳性精子的百分比和完整顶体的百分比。此外，分析精浆中丙二醛（MDA）的含量，并测量精浆中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）的水平。体外受精后，分析不同组的胚胎卵裂率和发育率，以比较精子的发育能力。结果表明，BAPT60 组解冻后精子活力明显高于 BAPT20、BAPT40 和对照组（P<0.05）。活精子的百分比从 BAPT20 组的 48.12±2.35%、BAPT40 组的 55.43±2.16%显著增加到 BAPT60 组的 57.53±3.15%。BAPT60 组 HOST 阳性精子的百分比明显高于 BAPT20、BAPT40 和对照组（P<0.05）。添加 BAPT 后，精浆和精子中的 MDA 水平显著降低（P<0.05）。此外，BAPT 实验组的 SOD、CAT 和 GSH-Px 水平明显高于对照组，这进一步表明 BAPT 显著抑制了绵羊精液冷冻过程中的活性氧（ROS）的产生。此外，BAPT40 和 BAPT60 组的胚胎卵裂率和发育率明显高于 BAPT20 和对照组（P<0.05）。综上所述，BAPT 通过抑制冷冻过程中的 ROS 产生来提高绵羊精子质量，可以作为绵羊精液冷冻保存的一种有前途的补充剂。
