Boar sperm are susceptible to oxidative damage caused by reactive oxygen species (ROS) during storage. Adenosine monophosphate (AMP)‐activated protein kinase (AMPK) is an important therapeutic target, because it is a cellular metabolism energy sensor and key signalling kinase in spermatozoa. We evaluated the effects of rosmarinic acid (RA), an antioxidant, on boar sperm during liquid storage to determine whether it protects boar sperm via AMPK activation. Boar ejaculates were diluted with Modena extender with different concentrations of RA and stored at 17°C for 9 days. Sperm quality parameters, antioxidant capacity, energy metabolism, AMPK phosphorylation and fertility were analysed. Compared with the control, 40 μmol/L significantly improved sperm motility, plasma membrane integrity and acrosome integrity (p < .05). The effective storage time of boar sperm was up to 9 days. On the third and seventh days, the sperm with RA exhibited increased total antioxidant capacity (T‐AOC), superoxide dismutase (SOD) activity, adenosine triphosphate (ATP) content, mitochondrial membrane potential (ΔΨm) and glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) activity, whereas malondialdehyde (MDA) content was significantly decreased (p < .05). Western blot showed that RA, as well as AICAR (AMPK activator), promoted AMPK phosphorylation, whereas Compound C (AMPK inhibitor) inhibited this effect. The sperm–zona pellucida binding experiment showed that 40 μmol/L RA increased the number of sperm attached to the zona pellucida (p < .05). These findings suggest meaningful methods for improved preservation of boar sperm in vitro and provide new insights into the mechanism by which RA protects sperm cells from oxidative damage via AMPK activation.

中文翻译：

---

迷迭香酸通过 AMPK 激活提高 17°C 下公猪精子质量、抗氧化能力和能量代谢。


公猪精子在储存过程中容易受到活性氧（ROS）引起的氧化损伤。单磷酸腺苷（AMP）激活蛋白激酶（AMPK）是一个重要的治疗靶点，因为它是精子中的细胞代谢能量传感器和关键信号激酶。我们评估了迷迭香酸 (RA)（一种抗氧化剂）在液体储存过程中对公猪精子的影响，以确定它是否通过 AMPK 激活来保护公猪精子。公猪精液用含有不同浓度 RA 的摩德纳稀释剂稀释，并在 17°C 下保存 9 天。分析精子质量参数、抗氧化能力、能量代谢、AMPK 磷酸化和生育力。与对照相比，40 μmol/L 显着改善精子活力、质膜完整性和顶体完整性（ p < .05）。公猪精子的有效保存时间长达9天。在第三天和第七天，患有 RA 的精子表现出总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）活性、三磷酸腺苷（ATP）含量、线粒体膜电位（ΔΨm）和甘油醛-3-磷酸脱氢酶（ GAPDH) 活性，而丙二醛 (MDA) 含量显着降低 ( p < .05)。蛋白质印迹显示 RA 以及 AICAR（AMPK 激活剂）促进 AMPK 磷酸化，而化合物 C（AMPK 抑制剂）抑制这种作用。精子与透明带结合实验表明，40 μmol/L RA 增加了附着在透明带上的精子数量（ p < .05）。这些发现提出了改善公猪精子体外保存的有意义的方法，并为 RA 通过 AMPK 激活保护精子细胞免受氧化损伤的机制提供了新的见解。