Lampiao Fanuel, du Plessis Stefan S
Department of Biomedical Sciences, Division of Medical Physiology, University of Stellenbosch, Tygerberg 7505, South Africa.
Asian J Androl. 2008 Sep;10(5):799-807. doi: 10.1111/j.1745-7262.2008.00421.x.
To investigate the in vitro effects of insulin and leptin on human sperm motility, viability, acrosome reaction and nitric oxide (NO) production.
Washed human spermatozoa from normozoospermic donors were treated with insulin (10 microIU) and leptin (10 nmol). Insulin and leptin effects were blocked by inhibition of their intracellular effector, phosphotidylinositol 3-kinase (PI3K), by wortmannin (10 micromol) 30 min prior to insulin and leptin being given. Computer-assisted semen analysis was used to assess motility after 1, 2 and 3 h of incubation. Viability was assessed by fluorescence-activated cell sorting using propidium iodide as a fluorescent probe. Acrosome-reacted cells were observed under a fluorescent microscope using fluorescein-isothiocyanate-Pisum sativum agglutinin as a probe. NO was measured after treating the sperm with 4,5-diaminofluorescein-2/diacetate (DAF-2/DA) and analyzed by fluorescence-activated cell sorting.
Insulin and leptin significantly increased total motility, progressive motility and acrosome reaction, as well as NO production.
This study showed the in vitro beneficial effects of insulin and leptin on human sperm function. These hormones could play a role in enhancing the fertilization capacity of human spermatozoa.
研究胰岛素和瘦素对人精子活力、生存力、顶体反应及一氧化氮(NO)生成的体外作用。
用来自正常精子捐献者的洗涤后人类精子,分别用胰岛素(10微国际单位)和瘦素(10纳摩尔)处理。在给予胰岛素和瘦素前30分钟,用渥曼青霉素(10微摩尔)抑制其细胞内效应物磷脂酰肌醇3激酶(PI3K),以阻断胰岛素和瘦素的作用。孵育1、2和3小时后,采用计算机辅助精液分析评估精子活力。用碘化丙啶作为荧光探针,通过荧光激活细胞分选评估精子生存力。用异硫氰酸荧光素-豌豆凝集素作为探针,在荧光显微镜下观察顶体反应的细胞。用4,5-二氨基荧光素-2/二乙酸酯(DAF-2/DA)处理精子后测量NO,并通过荧光激活细胞分选进行分析。
胰岛素和瘦素显著提高了总活力、前向运动能力、顶体反应以及NO的生成。
本研究显示了胰岛素和瘦素对人精子功能的体外有益作用。这些激素可能在增强人类精子的受精能力方面发挥作用。