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：

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迷迭香酸可通过AMPK活化改善公猪的精子质量，抗氧化能力和17°C的能量代谢。

公猪精子在储存过程中易受活性氧（ROS）引起的氧化损伤。单磷酸腺苷（AMP）活化的蛋白激酶（AMPK）是重要的治疗靶标，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）在储液期间对公猪精子的影响，以确定其是否通过AMPK活化保护公猪精子。将公猪射精用具有不同RA浓度的Modena增量剂稀释，并在17°C下保存9天。分析了精子质量参数，抗氧化能力，能量代谢，AMPK磷酸化和受精能力。与对照组相比，40μmol/ L显着改善了精子活力，质膜完整性和顶体完整性（p <.05）。公猪精子的有效储存时间长达9天。在第3天和第7天，RA精子的总抗氧化能力（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）。Western印迹显示RA和AICAR（AMPK激活剂）均促进AMPK磷酸化，而化合物C（AMPK抑制剂）抑制该作用。精子-透明带结合实验表明，40μmol/ L RA可增加附着于透明带的精子数量（p <.05）。这些发现为改善公猪精子的体外保存提供了有意义的方法，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见解。