基础医学与临床 ›› 2024, Vol. 44 ›› Issue (7): 940-946.doi: 10.16352/j.issn.1001-6325.2024.07.0940

• 研究论文 • 上一篇    下一篇

丙二酸二钠通过抑制琥珀酸脱氢酶活性损伤人精子运动功能

彭真1, 闻琴1, 卢京2, 涂泽梁2, 程一民2,3*   

  1. 1.宜春市人民医院 药学部,江西 宜春 336000;
    2.宜春学院 医学院 转化医学中心,江西 宜春 336000;
    3.广州中医药大学 分子节律与代谢研究所,广东 广州 510006
  • 收稿日期:2023-12-29 修回日期:2024-03-27 出版日期:2024-07-05 发布日期:2024-06-26
  • 通讯作者: *cym1169331029@outlook.com
  • 基金资助:
    国家自然科学基金 (82101690);江西省自然科学基金 (20224BAB216029);江西省教育厅科技项目 (GJJ2201749);宜春市指导性科技计划项目(2023ZDJH2699)

Disodium malonate impairs human sperm motility by inhibiting succinate dehydrogenase activity

PENG Zhen1, WEN Qin1, LU Jing2, TU Zeliang2, CHENG Yimin2,3*   

  1. 1. Department of Pharmacy, Yichun People's Hospital, Yichun 336000;
    2. Center for Translational Medicine, Department of Medicine, Yichun University, Yichun 336000;
    3. Institute of Molecular Rhythm and Metabolism, Guangzhou University of Chinese Medicine, Guangzhou 510006, China
  • Received:2023-12-29 Revised:2024-03-27 Online:2024-07-05 Published:2024-06-26
  • Contact: *cym1169331029@outlook.com

摘要: 目的 揭示琥珀酸脱氢酶(SDH)在人精子功能调控中的作用。方法 将分离纯化后的人精子样本与不同浓度(10、20、40 mmol/L)SDH抑制剂丙二酸二钠共孵育不同时间(1、2 h)后,试剂盒检测对照组及丙二酸二钠处理组SDH酶活性,Western blot检测SDH催化亚基(SDHA)蛋白水平。精子功能研究实验:1)通过计算机辅助精液分析系统检测丙二酸二钠对未获能精子重要运动参数[前向运动率(PR)、总活力(TM)、平均路径速度(VAP)]以及获能精子穿透粘性介质能力的影响;2)伊红-苯胺黑法评估丙二酸二钠对精子存活率的作用;3)PSA-FITC染色法检测丙二酸二钠处理对获能精子顶体反应发生率的影响。结果 丙二酸二钠不改变人精子中琥珀酸脱氢酶A(SDHA)蛋白表达水平,但其能够抑制SDH酶催化活性、精子前向运动率、总活力以及获能精子穿透粘性介质的能力,且上述抑制效应同丙二酸二钠浓度正相关。此外,丙二酸二钠不影响获能精子自发顶体反应发生率。结论 丙二酸二钠通过抑制SDH活性损伤人精子运动功能。

关键词: 琥珀酸脱氢酶, 丙二酸二钠, 精子功能, 精子运动

Abstract: Objective To investigate the impact of succinate dehydrogenase (SDH) on the modulation of human sperm functions. Methods The isolated human sperm were co-incubated with different concentrations (10, 20, 40 mmol/L) of SDH inhibitor disodium malonate for one or two hours. The activity of the SDH was measured by commercially available reagent kit, while the protein level of the SDH catalytic subunit SDHA was determined through Western blot analysis. Sperm functions were analyzed:1)The impact of disodium malonate on important motility parameters of un-capcitated sperm including progressive motility rate (PR), total motility (TM), average pathvelocity (VAP)and the ability of capacitated sperm to penetrate viscous media were be assessed using a computer aided semen analysis system. 2) Effect of disodium malonate on sperm survival rate was evaluated using the Eosin-Nigrosin microscopy. 3) The incidence of acrosome reaction in capacitated sperm was be detected by PSA-FITC staining assay following disodium malonate treatment. Results Disodium malonate had no effect on expression of SDH catalytic subunit SHDA protein in human sperm. However, it inhibited the catalytic activity of the SDH, sperm forward motility, total motility, and the ability of sperm to penetrate viscous media. These inhibitory effects were positively correlated with the concentration of disodium malonate. Furthermore, disodium malonate had no any influence on the occurrence of spontaneous acrosome reaction in capacitated sperm. Conclusions Disodium malonate impairs human sperm motility by inhibiting succinate dehydrogenase activity.

Key words: succinate dehydrogenase, disodium malonate, sperm function, sperm motility

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