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17β-estradiol modifies human spermatozoa mitochondrial function in vitro

机译:17β-雌二醇体外修饰人精子线粒体功能

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Background It is assumed that spermatozoa are target cells for estrogens however, the mechanism of their action is not fully understood. The aim of this study was to investigate the influence of 17β-estradiol (E2) on the human spermatozoa mitochondrial function. Methods The effects on spermatozoa of E2 at final concentrations of 10?10, 10?8 and 10?6 M were studied regarding the following phenomena: (1) kinetics of intracellular free calcium ions changes (using Fluo-3), (2) mitochondrial membrane potential ΔΨm (using JC-1 fluorochrome), (3) production of superoxide anion in mitochondria (using MitoSOX RED dye), (4) spermatozoa vitality (propidium iodide staining) and (5) phosphatidylserine membrane translocation (staining with annexin V marked with fluorescein). Results E2 initiated rapid (within a few seconds) dose dependent increase of intracellular free calcium ions concentration. E2 was changing the mitochondrial membrane potential: 10?8 M initiated significant increase of percentage of high ΔΨm spermatozoa while the 10?6 M induced significant decrease of high ΔΨm cells. In spermatozoa stimulated with E2 10?6 M a significant increase of mitochondrial superoxide anion level was observed. 2?h incubation of spermatozoa with E2 did not alter cells vitality nor stimulated phosphatidylserine membrane translocation, for all three doses. Conclusions 17β-estradiol affected the human spermatozoa mitochondrial function. E2 in low concentration improved while in high concentration might deteriorate mitochondrial function.
机译:背景技术假定精子是雌激素的靶细胞,但是,其作用机理尚不完全清楚。这项研究的目的是调查17β-雌二醇(E2)对人精子线粒体功能的影响。方法研究以下现象对终浓度为10 ?10 ,10 ?8 和10 ?6 M的E2精子的影响:(1)细胞内游离钙离子变化的动力学(使用Fluo-3),(2)线粒体膜电位ΔΨ m (使用JC-1荧光染料),(3)线粒体中超氧阴离子的产生(使用MitoSOX RED染料),(4)精子活力(碘化丙啶染色)和(5)磷脂酰丝氨酸膜易位(用标记有荧光素的膜联蛋白V染色)。结果E2启动(在几秒钟内)剂量依赖性细胞内游离钙离子浓度的快速增加。 E2正在改变线粒体膜电位:10 ?8 M引起高ΔΨ m 精子的百分比显着增加,而10 ?6 M引起高ΔΨm细胞明显减少。在E2 10 ?6 M刺激下的精子中,线粒体超氧阴离子水平显着增加。对于所有三种剂量,用E2孵育2?h都不会改变细胞活力,也不会刺激磷脂酰丝氨酸膜移位。结论17β-雌二醇影响人精子线粒体功能。低浓度的E2有所改善,而高浓度的E2可能会使线粒体功能恶化。

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