Zadeh Hashem Elham, Eslami Mohsen
Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
Department of Theriogenology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.
Cell Tissue Bank. 2018 Mar;19(1):97-111. doi: 10.1007/s10561-016-9604-3. Epub 2016 Nov 25.
The present experiment was conducted to evaluate the effect of kinetin on ram semen quality during cold storage. Ejaculates were collected using an artificial vagina from five Qezel rams. Ejaculates which met the criteria (volume of 0.75-2 ml; minimum spermatozoa concentration of 2.5 × 109 spermatozoa/ml and forward progressive motility of 80%), were pooled, diluted with extender without kinetin (control) or enriched with 25 (K 25), 50 (K 50), 100 (K 100) and 200 (K 200) μM kinetin at a final concentration of 500 × 106 spermatozoa per mL. Spermatozoa motion characteristics were evaluated by computer-assisted sperm analysis. In addition, percent of viability (spermatozoa showing no color was considered to be alive) and spermatozoa with intact plasma membrane (spermatozoa with curled/swollen tail was considered healthy) were determined. Moreover, amounts of malondialdehyde (MDA), total antioxidant activity (AOA), nitric oxide (NO) and superoxide dismutase (SOD) activity were determined in the seminal plasma and spermatozoa at 0, 24, 48 and 72 h of storage. Higher percent of total and forward progressive motility was observed in K 25, K 50 and K 100 groups compared to control group at 72 h of storage (P < 0.001). Moreover, K 25 (78.61 ± 1.11%), K 50 (82.46 ± 1.08%) and K 100 (82.96 ± 1.49%) groups showed higher percentage of viability compared to control (72.38 ± 1.49%) at 72 h of storage (P < 0.05). Semen enrichment with kinetin resulted in the higher percent of intact plasma membrane of spermatozoa at 48 and 72 h (P < 0.001). Amounts of MDA were lower and amounts of AOA were higher in K 50 and K 100 groups compared to control at 48 and 72 h (P < 0.05). There were no significant differences in NO levels and SOD activities of seminal plasma and spermatozoa among groups during the experiment. The present experiment revealed that kinetin at proper concentration could enhances spermatozoa kinematics, viability, spermatozoa plasma membrane functionality and amounts of AOA and reduces the level of lipid peroxidation during chilled storage of ram semen.
本实验旨在评估激动素对冷藏期间公羊精液品质的影响。使用人工阴道从五只盖泽尔公羊采集精液。将符合标准(体积为0.75 - 2毫升;精子最低浓度为2.5×10⁹个精子/毫升且前向运动性为80%)的精液汇集,用不含激动素的稀释液(对照组)或添加25(K 25)、50(K 50)、100(K 100)和200(K 200)μM激动素的稀释液进行稀释,最终浓度为每毫升500×10⁶个精子。通过计算机辅助精子分析评估精子运动特征。此外,测定活力百分比(无颜色的精子被视为存活)和质膜完整的精子(尾巴卷曲/肿胀的精子被视为健康)。此外,在储存0、24、48和72小时时,测定精浆和精子中丙二醛(MDA)、总抗氧化活性(AOA)、一氧化氮(NO)和超氧化物歧化酶(SOD)活性的含量。在储存72小时时,与对照组相比,K 25、K 50和K 100组的总运动性和前向运动性百分比更高(P < 0.001)。此外,在储存72小时时,K 25(78.61±1.11%)、K 50(82.46±1.08%)和K 100(82.96±1.49%)组的活力百分比高于对照组(72.38±1.49%)(P < 0.05)。用激动素富集精液导致在48和72小时时精子质膜完整的百分比更高(P < 0.001)。在48和72小时时,与对照组相比,K 50和K 100组的MDA含量较低,AOA含量较高(P < 0.05)。在实验期间,各组精浆和精子的NO水平和SOD活性没有显著差异。本实验表明,适当浓度的激动素可增强公羊精液冷藏期间精子的运动学、活力、精子质膜功能以及AOA含量,并降低脂质过氧化水平。