CONTEXT Perfluoroalkyl-substances (PFAS) are chemical additives considered harmful for humans. We recently showed that accumulation of perfluoro-octanoic acid (PFOA) in human semen of exposed subjects was associated with altered motility parameters of sperm cells, suggesting direct toxicity. OBJECTIVES To determine whether direct exposure of human spermatozoa to PFOA was associated to impairment of cell function. PATIENTS AND METHODS Spermatozoa isolated from semen samples of ten normozoospermic healthy donors were exposed up to 2 h to PFOA, at concentrations from 0.1 to 10 ng/mL. Viability and motility parameters were evaluated by Sperm Class Analyser. Cell respiratory function was assessed by both mitochondrial probe JC-1 and respiratory control ratio (RCR) determination. Sperm accumulation of PFOA was quantified by liquid chromatography-mass spectrometry. Expression of organic ion-transporters OATP1 and SLCO1B2 was assessed by immunofluorescence and respective role in PFOA accumulation was evaluated by either blockade with probenecid or membrane scavenging through β-cyclodextrin (β-CD). Plasma membrane fluidity and electrochemical potential (ΔΨp) were evaluated, respectively, with Merocyanine-540 and Di-3-ANEPPDHQ fluorescent probes. RESULTS Compared to untreated controls, a threefold increase of the percentage of non-motile sperms was observed after 2 h of exposure to PFOA regardless of the concentration of PFOA, whilst RCR was significantly reduced. Only scavenging with β-CD was effective in reducing PFOA accumulation, suggesting membrane involvement. Altered membrane fluidity, reduced ΔΨp and sperm motility loss associated with exposure to PFOA were reverted by β-CD treatment. CONCLUSION PFOA alters human sperm motility through plasma-membrane disruption, an effect recovered by incubation with β-CD.

中文翻译：

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全氟辛酸会通过质膜的改变损害精子的活力。

背景全氟烷基物质（PFAS）是被认为对人体有害的化学添加剂。我们最近显示，全氟辛酸（PFOA）在暴露对象的人类精液中积累与精子细胞的运动参数改变有关，提示直接毒性。目的确定人类精子直接暴露于PFOA是否与细胞功能受损有关。患者和方法从十个正常精子健康供体的精液样本中分离出的精子暴露于PFOA达2 h，浓度为0.1至10 ng / mL。通过Sperm Class Analyser评估生存力和运动性参数。通过线粒体探针JC-1和呼吸控制率（RCR）测定来评估细胞的呼吸功能。通过液相色谱-质谱法定量分析PFOA的精子积累。通过免疫荧光评估有机离子转运蛋白OATP1和SLCO1B2的表达，并通过丙磺舒的阻滞或通过β-环糊精（β-CD）清除膜来评估PFOA积累中的各自作用。使用Merocyanine-540和Di-3-ANEPPDHQ荧光探针分别评估了质膜的流动性和电化学势（ΔΨp）。结果与未治疗的对照组相比，暴露于PFOA 2 h后，非活动精子的百分比增加了三倍，而与PFOA的浓度无关，而RCR却显着降低。仅用β-CD清除可有效减少PFOA的积累，提示膜参与。膜流动性改变，β-CD治疗可逆转因暴露于PFOA而引起的Δp降低和精子活力丧失。结论PFOA通过破坏血浆膜而改变人的精子运动能力，这种作用可以通过与β-CD一起温育来恢复。