Department of Zoology, The University of Buner, Khyber Pakhtunkhwa (KP), Pakistan; Reproductive Physiology Laboratory, Department of Animal Sciences, Quaid-i-Azam University (QAU), Islamabad, Pakistan.
Reproductive Physiology Laboratory, Department of Animal Sciences, Quaid-i-Azam University (QAU), Islamabad, Pakistan.
Cryobiology. 2020 Dec;97:101-109. doi: 10.1016/j.cryobiol.2020.09.010. Epub 2020 Sep 30.
The disparity between the endogenous antioxidants concentration and free radicals in spermatozoa results in reactive oxygen species (ROS) generation. In this prospect, epigallocatechin-3-gallate (EGCG) preserves vigorous antioxidant features. Current study explored the influence of EGCG in a cryo-diluent media on microscopic parameters, oxidative stress parameters, and fertility potential of buffalo spermatozoa during cryopreservation. Concisely, collected semen from three donor bulls for four times were then evaluated for volume, motility, concentrations and then dilution in a cryo-diluent media with different concentrations of EGCG (EGCG-0 = control; EGCG-50 = 50 μM, EGCG-100 = 100 μM, EGCG-200 = 200 μM, and EGCG-300 = 300 μM) at 37 °C, cooled to 4 °C in 2 h, equilibrated for 4 h at 4 °C, and cryopreserved. At post-thawing, Computer-Assisted Sperm motion Analysis motilities (total and progressive, %) and rapid velocity (%), plasma membrane functionality, supravital plasma membrane integrity, and mitochondrial potential (%) were found higher (P < 0.05) in EGCG-200, and EGCG-300 than control, whereas average-path, straight-line, and curved-linear velocities (μm/sec), and acrosome integrity (%) were recorded higher in EGCG-300 than control. Further, comet length (μm), and tail length (μm), LPO (lipid peroxidation, μM/mL), and apoptosis-like changes (%) in spermatozoa were significantly decreased in EGCG-300 than control. Seminal plasma antioxidant enzymes activities (glutathione peroxidase, U/mL, and superoxide dismutase, U/mL) were increased with EGCG-300 than control. Moreover, EGCG-300 addition in a cryo-diluent media improves the fertility potential (%) of buffalo spermatozoa. In a nutshell, the inclusion of EGCG-300 in a cryo-diluent media enhances post-thaw microscopic parameters, and fertility potential, whereas decreases oxidative stress parameters in buffalo spermatozoa.
内源性抗氧化剂浓度与精子中自由基之间的差异导致活性氧（ROS）的产生。在这种情况下，表没食子儿茶素没食子酸酯（EGCG）具有强大的抗氧化特性。本研究探讨了 EGCG 在冷冻稀释液中的浓度对水牛精子冷冻保存过程中微观参数、氧化应激参数和生育能力的影响。简而言之，从三头公牛收集精液四次，然后评估其体积、活力、浓度，然后在含有不同浓度 EGCG（EGCG-0=对照；EGCG-50=50 μM、EGCG-100=100 μM、EGCG-200=200 μM 和 EGCG-300=300 μM）的冷冻稀释液中稀释，在 37°C 下冷却至 4°C 2 小时，在 4°C 下平衡 4 小时，然后冷冻保存。在解冻后，计算机辅助精子运动分析发现，总运动和渐进运动（%）和快速运动（%）、质膜功能、超活质膜完整性和线粒体潜能（%）在 EGCG-200 和 EGCG-300 中均高于对照组，而平均路径、直线和曲线速度（μm/sec）和顶体完整性（%）在 EGCG-300 中均高于对照组。此外，在 EGCG-300 中，精子的彗星长度（μm）和尾部长度（μm）、脂质过氧化（LPO，μM/mL）和凋亡样变化（%）均显著低于对照组。在 EGCG-300 中，精子中的谷胱甘肽过氧化物酶（U/mL）和超氧化物歧化酶（U/mL）等抗氧化酶活性增加。此外，在冷冻稀释液中添加 EGCG-300 可提高水牛精子的生育能力（%）。简而言之，在冷冻稀释液中添加 EGCG-300 可提高水牛精子解冻后的微观参数和生育能力，同时降低精子的氧化应激参数。
