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李榕楠, 朱振东, 郑以, 吕英华, 田秀娥, 吴德, 王永军, 曾文先. 2020: 二甲双胍调控AMPK介导的能量代谢改善体外保存猪精子品质. 动物学研究, 41(5): 527-538. DOI: 10.24272/j.issn.2095-8137.2020.074
引用本文: 李榕楠, 朱振东, 郑以, 吕英华, 田秀娥, 吴德, 王永军, 曾文先. 2020: 二甲双胍调控AMPK介导的能量代谢改善体外保存猪精子品质. 动物学研究, 41(5): 527-538. DOI: 10.24272/j.issn.2095-8137.2020.074
Rong-Nan Li, Zhen-Dong Zhu, Yi Zheng, Ying-Hua Lv, Xiu-E Tian, De Wu, Yong-Jun Wang, Wen-Xian Zeng. 2020: Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro. Zoological Research, 41(5): 527-538. DOI: 10.24272/j.issn.2095-8137.2020.074
Citation: Rong-Nan Li, Zhen-Dong Zhu, Yi Zheng, Ying-Hua Lv, Xiu-E Tian, De Wu, Yong-Jun Wang, Wen-Xian Zeng. 2020: Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro. Zoological Research, 41(5): 527-538. DOI: 10.24272/j.issn.2095-8137.2020.074

二甲双胍调控AMPK介导的能量代谢改善体外保存猪精子品质

Metformin improves boar sperm quality via 5′-AMP-activated protein kinase-mediated energy metabolism in vitro

  • 摘要: 精子的运动、获能及顶体反应等过程需要大量的能量。维持精子能量稳态对于改善猪精子体外保存的效果至关重要。二甲双胍通过激活AMP依赖的蛋白激酶(AMPK)维持细胞内能量代谢的稳态。因此,本研究旨在探究二甲双胍能否通过AMPK调控能量代谢而改善猪精子的品质。本研究对猪精子活力参数、质膜完整性、线粒体膜电位、ATP含量、葡萄糖摄取量和乳酸外排量等指标进行检测,并运用蛋白质免疫印迹和免疫荧光技术检测AMPK与磷酸化AMPK(172位点;p-AMPK)在精子中的定位和表达水平。结果显示,在17 °C保存过程中,添加二甲双胍显著提高精子活力、线粒体膜电位和ATP含量。同时,发现AMPK定位于精子的顶体、头尾连接段和中段,且p-AMPK分布于顶体后区、头尾连接段和中段。此外,添加二甲双胍显著提高37 °C孵育4h后的精子活力、线粒体膜电位、ATP含量、p-AMPK水平、葡萄糖摄取量和乳酸外排量。然而,添加AMPK抑制剂Compound C会抑制二甲双胍对精子的作用。综上所述,二甲双胍能够激活AMPK,进而维持猪精子能量代谢和线粒体的活性,从而改善精子功能与精液保存的效果。

     

    Abstract: Sperm are specialized cells that require adenosine triphosphate (ATP) to support their function. Maintaining sperm energy homeostasis in vitro is vitally important to improve the efficacy of boar sperm preservation. Metformin can activate 5′-AMP-activated protein kinase (AMPK) to improve metabolic flexibility and maintain energy homeostasis. Thus, the aim of the present study was to investigate whether metformin can improve boar sperm quality through AMPK mediation of energy metabolism. Sperm motility parameters, membrane integrity, acrosome integrity, mitochondrial membrane potential (ΔΨm), ATP content, glucose uptake, and lactate efflux were analyzed. Localization and expression levels of AMPK and phospho-Thr172-AMPK (p-AMPK) were also detected by immunofluorescence and western blotting. We found that metformin treatment significantly increased sperm motility parameters, ΔΨm, and ATP content during storage at 17 °C. Moreover, results showed that AMPK was localized at the acrosomal region, connecting piece, and midpiece of sperm and p-AMPK was distributed at the post-acrosomal region, connecting piece, and midpiece. When sperm were incubated with metformin for 4 h at 37 °C, sperm motility parameters, ΔΨm, ATP content, p-AMPK, glucose uptake, and lactate efflux all significantly increased, whereas the addition of Compound C treatment, an inhibitor of AMPK, counteracted these positive effects. Together, our results suggest that metformin promotes AMPK activation, which contributes to the maintenance of energy hemostasis and mitochondrial activity, thereby maintaining boar sperm functionality and improving the efficacy of semen preservation.

     

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