抗氧化劑添加於豬精液冷凍保存之影響

Effect of antioxidants supplementation on boar semen cryopreservation

章嘉潔(Chia-Chieh Chang) ； 吳昇陽(Sheng-Yang Wu)

《畜產研究》 52卷2期 (2019/06) Pp. 72-80

摘要

本試驗探討三種抗氧化劑α-生育醇（α-tocopherol）、丁羥基甲苯（butylated hydroxytoluene, BHT）及麩胱甘肽（glutathione, GSH），添加於豬冷凍稀釋液對解凍後精液品質之影響。收集5頭杜洛克公豬之新鮮精液，以Lactose egg yolk（LEY）之冷凍精液稀釋液稀釋，精子最終稀釋濃度為5 × 10^8 cells /mL。試驗一之分組分為對照組（LEY組）、添加5 mM GSH、1 mM BHT及200 μg/mL α-tocopherol入冷凍稀釋液等4組，以評估解凍後精子活力、精子前進式活力、精子活力各項移動參數，及精子頭帽完整性等項目之差異。結果顯示精液解凍後體外培養2至6小時，5 mM GSH添加組精子活力及精子快速前進式活力均較200 μg/mL α-tocopherol添加組差（P ＜ 0.05）。200 μg/mLα-tocopherol或1 mM BHT添加組與對照組進行比較，統計無顯著差異。在精子活力各項移動參數方面，精液解凍後培養5分鐘，5 mM GSH添加組精子活力移動參數VAP、VSL及VCL均較1 mM BHT組呈顯著低之現象（P ＜ 0.05）。添加濃度1 mM BHT或200 μg/mL α-tocopherol於解凍後精子頭帽完整率，與對照組或5 mM GSH添加組比較，結果呈顯著改善（P ＜ 0.05）。試驗二之分組分為對照組（LEY組）、添加300、400 或500 μg/mL α-tocopherol濃度加入冷凍稀釋液等4組，評估解凍後的精子活力、精子前進式活力及精子活力各項移動參數，結果顯示精子活力及精子快速前進式活力及精子活力等各項移動參數，於對照組與不同濃度α-tocopherol添加組間均無顯著差異。本研究冷凍稀釋液中添加5 mM GSH、1 mM BHT及200、300、400與500 μg/mL α-tocopherol組之解凍後豬精子的相關活力參數並未見顯著改善效果。

關鍵字

豬 ； 冷凍精液 ； 抗氧化劑

並列摘要

The aim of this study was to investigate the effects of three antioxidants α-tocopherol, butlated hydroxytoluene (BHT) and glutathione (GSH) were added to freezing extender on the quality of frozen-thawed boar semen. Semen collected from five Duroc boars were diluted with Lactose-egg yolk (LEY) extender as control group, which it brought to 5 × 10^8 cell/mL in the final concentration. In the first experiment, four groups were divided with adding 5 mM GSH, 1mM BHT and 200 μg/mL α-tocopherol into freezing extenders. The percentage of sperm motility, rapid progressive motility, motility kinetic variables parameters and acrosome integrity were evaluated. The results showed that the percentage of total motility and rapid progressive motility in 5mM GSH supplemented freezing extender after thawing for 2-6 hrs were lower than in 200 μg/mL α-tocopherol group (P ＜0.05). No significant differences in the percentage of total motility and rapid progressive motility were observed either between control group and 200 μg/mL α-tocopherol or 1mM BHT supplemented. The results showed that the percentage of sperm motion parameters (VAP, VSL and VCL) of semen cryopreserved with freezing extender after thawing for 5min were lower in 5 mM GSH group than in the 1mM BHT group (P ＜ 0.05). Acrosome integrity results demonstrated that the intact acrosome was significantly higher in the 1mM BHT or 200 μg/mL α-tocopherol group than that in the control or than that in 5 mM GSH group (P ＜ 0.05). In the second experiment, the experimental designs are separated into 4 groups, inclusive of control group, and the 300, 400 or 500 μg/mL α-tocopherol supplementation group in the boar semen extender during cryopreservation on post-thawed sperm motility, rapid progressive motility and motility kinetic variables parameters. There is no significant difference between the α-tocopherol supplemented and control group. In conclusion, the addition of 5 mM GSH, 1 mM BHT and 200, 300, 400 or 500 μg/mL α-tocopherol to the freezing extender demonstrate no any improvement in sperm motility parameters.

並列關鍵字

Boar ； Frozen Semen ； Antioxidant