Department of Growth and Reproduction, Copenhagen University Hospital, Rigshospitalet, Copenhagen, Denmark.
International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC), Rigshospitalet, University of Copenhagen, Copenhagen, Denmark.
Front Endocrinol (Lausanne). 2020 May 19;11:324. doi: 10.3389/fendo.2020.00324. eCollection 2020.
Evidence suggests that bisphenol A diglycidyl ether (BADGE), bisphenol A (BPA), and BPA analogs can interfere with human male fertility. However, the effect directly on human sperm function is not known. The CatSper Ca channel in human sperm controls important sperm functions and is necessary for normal male fertility. Environmental chemicals have been shown to activate CatSper and thereby affect Ca signaling in human sperm. BPA has previously been investigated for effects on Ca signaling human sperm, whereas the effects of other BPA analogs are currently unknown. The aim of this study is thus to characterize the effect of BADGE, BPA, and the eight analogs BPG, BPAF, BPC, BPB, BPBP, BPE, BPF, BPS on Ca signaling, and CatSper in human sperm. Direct effects of the bisphenols on Ca signaling in human sperm cells were evaluated using a Ca fluorimetric assay measuring changes in intracellular Ca. Effects via CatSper were assessed using the specific CatSper inhibitor RU1968. Effects on human sperm function was assessed using an image cytometry-based acrosome reaction assay and the modified Kremer's sperm-mucus penetration assay. At 10 μM the bisphenols BPG, BPAF, BPC, BADGE, BPB, and BPBP induced Ca signals in human sperm cells, whereas BPE, BPF, BPS, and BPA had no effect. The efficacy of the chemicals at 10 μM is BPG > BPAF > BPC > BADGE > BPB > BPBP. Dose-response relations of BPG, BPAF, BPC, BADGE, BPB, and BPBP yielded EC50-values in the nM-μM range. The induced Ca signals were almost completely abolished using the CatSper inhibitor RU1968, indicating an effect of the bisphenols on CatSper. All bisphenols, except BPBP, were found to dose-dependently inhibit progesterone-induced Ca signals, with BPG and BPAF displaying inhibition even in low μM doses. BPG and BPAF were shown to affect human sperm function in a progesterone-like manner. Our results show that the bisphenols BPG, BPAF, BPC, BADGE, BPB, and BPBP can affect Ca signaling in human sperm cells through activation of CatSper. This could potentially disrupt human sperm function by interfering with normal CatSper-signaling and thus be a contributing factor in human infertility, either alone or in mixtures with other chemicals.
有证据表明,双酚 A 二缩水甘油醚(BADGE)、双酚 A(BPA)和 BPA 类似物会干扰男性生育能力。然而,其对人类精子功能的直接影响尚不清楚。人类精子中的 CatSper Ca 通道控制着重要的精子功能,是正常男性生育能力所必需的。环境化学物质已被证明可以激活 CatSper,从而影响人类精子中的 Ca 信号转导。BPA 先前已被研究用于对人类精子 Ca 信号转导的影响,而其他 BPA 类似物的影响目前尚不清楚。因此,本研究旨在研究 BADGE、BPA 和 8 种类似物 BPG、BPAF、BPC、BPB、BPBP、BPE、BPF 和 BPS 对人类精子 Ca 信号转导和 CatSper 的影响。通过使用测量细胞内 Ca 变化的 Ca 荧光比色法评估双酚对人类精子细胞 Ca 信号转导的直接影响。通过 CatSper 特异性抑制剂 RU1968 评估通过 CatSper 的作用。使用基于图像细胞术的顶体反应测定法和改良的 Kremer 精子-黏液渗透测定法评估对人类精子功能的影响。在 10 μM 时,双酚 BPG、BPAF、BPC、BADGE、BPB 和 BPBP 诱导人类精子细胞产生 Ca 信号,而 BPE、BPF、BPS 和 BPA 则没有影响。在 10 μM 时,这些化学物质的功效为 BPG>BPAF>BPC>BADGE>BPB>BPBP。BPG、BPAF、BPC、BADGE、BPB 和 BPBP 的剂量反应关系产生的 EC50 值在纳摩尔-微摩尔范围内。使用 CatSper 抑制剂 RU1968 几乎完全消除了诱导的 Ca 信号,表明双酚对 CatSper 的作用。除 BPBP 外,所有双酚均被发现以剂量依赖的方式抑制孕酮诱导的 Ca 信号,BPG 和 BPAF 甚至在低 μM 剂量下就显示出抑制作用。BPG 和 BPAF 以类似于孕酮的方式影响人类精子功能。我们的研究结果表明,双酚 BPG、BPAF、BPC、BADGE、BPB 和 BPBP 可通过激活 CatSper 影响人类精子细胞中的 Ca 信号转导。这可能会通过干扰正常的 CatSper 信号转导而破坏人类精子功能,从而成为人类不育的一个促成因素,无论是单独存在还是与其他化学物质混合存在。