Abstract：
BACKGROUND: Various antioxidant substances are added to sperm extenders to protect spermatozoa against oxidative stress and cryodamage.
OBJECTIVE: To investigate the effects of the flavonoid diosmin (DIO) and a flavanone glycoside naringin (NAR) on the freezability of ram semen.
METHODS: In this study, six Merino rams were used during the breeding season. The ejaculates were pooled after collection from the rams. Pooled ejaculates were divided into six groups: control, NAR 1 mM, NAR 2 mM, NAR 4 mM, DIO 2 mM, and DIO 4 mM, and then diluted with a TRIS-based diluent. The pooled semen was equilibrated, placed in 0.25 mL pipettes with 10 × 10 7 sperm cells in each pipette, and frozen in liquid nitrogen vapor. After 24 h, the pipettes were thawed at 37 degree C for 25 s and analyzed in terms of spermatological parameters.
RESULTS: The highest plasma membrane integrity ratio was found in the DIO 4 mM group, whereas a statistically significant difference was found between the NAR 1 mM and NAR 2 mM groups (p < 0.05). While the DIO 4 mM group had the highest acrosome integrity rate, a statistically significant difference was found between the other groups (p < 0.05). Mitochondrial activity was the highest in the NAR 4 mM, DIO 4 mM and DIO 2 mM groups (p < 0.05). In the analysis of the sperm membrane lipid profile, it was observed that the DIO group had the highest lipid-phospholipid ratio. In sperm membrane protein profile analysis, it was found that both additives exerted protective effects at different levels. The highest total protein content was seen in the DIO 4 mM and NAR 4 mM groups. 8-hydroxydeoxyguanosine (8-OhDG) positivity was more common in the control group than in the DIO and NAR groups. Cu-Zn superoxide dismutase (SOD) expression was lower in the control group and more intense in all other groups. Positive results were especially observed in the acrosome of the sperm cells.
CONCLUSIONS: The addition of NAR and DIO to the ram semen extender increased the quality of sperm parameters after the freeze-thaw process. Doi.org/10.54680/fr24510110412.
OBJECTIVE: To investigate the effects of the flavonoid diosmin (DIO) and a flavanone glycoside naringin (NAR) on the freezability of ram semen.
METHODS: In this study, six Merino rams were used during the breeding season. The ejaculates were pooled after collection from the rams. Pooled ejaculates were divided into six groups: control, NAR 1 mM, NAR 2 mM, NAR 4 mM, DIO 2 mM, and DIO 4 mM, and then diluted with a TRIS-based diluent. The pooled semen was equilibrated, placed in 0.25 mL pipettes with 10 × 10 7 sperm cells in each pipette, and frozen in liquid nitrogen vapor. After 24 h, the pipettes were thawed at 37 degree C for 25 s and analyzed in terms of spermatological parameters.
RESULTS: The highest plasma membrane integrity ratio was found in the DIO 4 mM group, whereas a statistically significant difference was found between the NAR 1 mM and NAR 2 mM groups (p < 0.05). While the DIO 4 mM group had the highest acrosome integrity rate, a statistically significant difference was found between the other groups (p < 0.05). Mitochondrial activity was the highest in the NAR 4 mM, DIO 4 mM and DIO 2 mM groups (p < 0.05). In the analysis of the sperm membrane lipid profile, it was observed that the DIO group had the highest lipid-phospholipid ratio. In sperm membrane protein profile analysis, it was found that both additives exerted protective effects at different levels. The highest total protein content was seen in the DIO 4 mM and NAR 4 mM groups. 8-hydroxydeoxyguanosine (8-OhDG) positivity was more common in the control group than in the DIO and NAR groups. Cu-Zn superoxide dismutase (SOD) expression was lower in the control group and more intense in all other groups. Positive results were especially observed in the acrosome of the sperm cells.
CONCLUSIONS: The addition of NAR and DIO to the ram semen extender increased the quality of sperm parameters after the freeze-thaw process. Doi.org/10.54680/fr24510110412.
摘要：
背景：将各种抗氧化剂添加到精子补充剂中以保护精子免受氧化应激和冷冻损伤。
目的：研究黄酮类地奥司明（DIO）和黄烷酮苷柚皮苷（NAR）对公羊精液冷冻能力的影响。
方法：在本研究中，繁殖季节使用了六只Merino公羊。从公羊中收集后，将射精收集起来。集合射精分为六组：对照组，NAR1mM,NAR2mM，NAR4mM,DIO2mM,和DIO4mM，然后用基于TRIS的稀释剂稀释。汇集的精液被平衡，放置在0.25毫升移液管中，每个移液管中有10×107个精子细胞，在液氮蒸气中冷冻.24小时后，将移液管在37°C下解冻25s，并根据精子参数进行分析。
结果：在DIO4mM组中发现最高的质膜完整性比，而NAR1mM和NAR2mM组之间的差异有统计学意义(p<0.05)。虽然DIO4mM组的顶体完整性率最高，其他组间差异有统计学意义(p<0.05)。线粒体活性在NAR4mM中最高，DIO4mM和DIO2mM组(p<0.05)。在对精子膜脂质谱的分析中，观察到DIO组具有最高的脂质-磷脂比率。在精子膜蛋白谱分析中，发现两种添加剂在不同程度上发挥了保护作用。在DIO4mM和NAR4mM组中观察到最高的总蛋白质含量。8-羟基脱氧鸟苷（8-OhDG）阳性在对照组中比在DIO和NAR组中更常见。Cu-Zn超氧化物歧化酶（SOD）的表达在对照组中较低，而在所有其他组中更强烈。在精子细胞的顶体中尤其观察到阳性结果。
结论：将NAR和DIO添加到公羊精液补充剂中，提高了冻融过程后精子参数的质量。Doi.org/10.54680/fr24510110412。
目的：研究黄酮类地奥司明（DIO）和黄烷酮苷柚皮苷（NAR）对公羊精液冷冻能力的影响。
方法：在本研究中，繁殖季节使用了六只Merino公羊。从公羊中收集后，将射精收集起来。集合射精分为六组：对照组，NAR1mM,NAR2mM，NAR4mM,DIO2mM,和DIO4mM，然后用基于TRIS的稀释剂稀释。汇集的精液被平衡，放置在0.25毫升移液管中，每个移液管中有10×107个精子细胞，在液氮蒸气中冷冻.24小时后，将移液管在37°C下解冻25s，并根据精子参数进行分析。
结果：在DIO4mM组中发现最高的质膜完整性比，而NAR1mM和NAR2mM组之间的差异有统计学意义(p<0.05)。虽然DIO4mM组的顶体完整性率最高，其他组间差异有统计学意义(p<0.05)。线粒体活性在NAR4mM中最高，DIO4mM和DIO2mM组(p<0.05)。在对精子膜脂质谱的分析中，观察到DIO组具有最高的脂质-磷脂比率。在精子膜蛋白谱分析中，发现两种添加剂在不同程度上发挥了保护作用。在DIO4mM和NAR4mM组中观察到最高的总蛋白质含量。8-羟基脱氧鸟苷（8-OhDG）阳性在对照组中比在DIO和NAR组中更常见。Cu-Zn超氧化物歧化酶（SOD）的表达在对照组中较低，而在所有其他组中更强烈。在精子细胞的顶体中尤其观察到阳性结果。
结论：将NAR和DIO添加到公羊精液补充剂中，提高了冻融过程后精子参数的质量。Doi.org/10.54680/fr24510110412。
