通过对精子特异性和类固醇激活的 CatSper Ca 通道的作用，研究农药对人精子细胞钙信号转导的内分泌干扰作用的体外研究。

In vitro investigation of endocrine disrupting effects of pesticides on Ca-signaling in human sperm cells through actions on the sperm-specific and steroid-activated CatSper Ca-channel.

机构信息

Department of Growth and Reproduction, Copenhagen University Hospital, Rigshospitalet, Denmark; International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC), Rigshospitalet, University of Copenhagen, Denmark.

出版信息

Environ Int. 2022 Sep;167:107399. doi: 10.1016/j.envint.2022.107399. Epub 2022 Jul 9.

DOI:10.1016/j.envint.2022.107399

PMID:35853389

Abstract

BACKGROUND

Ca-signaling controls sperm cell functions necessary for successful fertilization. Multiple endocrine disrupting chemicals have been found to interfere with normal Ca-signaling in human sperm cells through an activation of the sperm-specific CatSper Ca-channel, which is vital for normal male fertility.

OBJECTIVES

We investigated 53 pesticides for their ability to interfere with CatSper mediated Ca-signaling and function in human sperm cells.

METHODS

Effects of the pesticides on Ca-signaling in human sperm cells were evaluated using a Ca-fluorometric assay. Effects via CatSper were assessed using the specific CatSper inhibitor RU1968. Effects on human sperm function and viability were assessed using an image cytometry-based acrosome reaction assay and the modified Kremer's sperm-mucus penetration assay.

RESULTS

28 of 53 pesticides were found to induce Ca-signals in human sperm cells at 10 µM. The majority of these 28 active pesticides induced Ca-signals through CatSper and interfered with subsequent Ca-signals induced by the two endogenous CatSper ligands progesterone and prostaglandin E. Multiple active pesticides were found to affect Ca-mediated sperm functions and viability at 10 µM. Low nM dose mixtures of the active pesticides alone or in combination with other environmental chemicals were found to significantly induce Ca-signals and inhibit Ca-signals induced subsequently by progesterone and prostaglandin E.

CONCLUSIONS

Our results show that pesticides, both alone and in low nM dose mixtures, interfere with normal Ca-signaling in human sperm cells in vitro in low nM concentrations. Biomonitoring of the active pesticides in relevant matrices such as blood and reproductive fluids is very limited and the effects of real time human pesticide exposure on human sperm cells and fertility thus remains largely unknown. To which extent human pesticide exposure affects the chances of a successful fertilization in humans in vivo needs further research.

摘要

背景

钙信号控制精子细胞的功能，这些功能对于成功受精是必要的。已经发现，多种内分泌干扰化学物质通过激活精子特异性 CatSper 钙通道来干扰人精子细胞中的正常钙信号，CatSper 钙通道对正常男性生育力至关重要。

目的

我们研究了 53 种农药干扰人精子细胞中 CatSper 介导的钙信号和功能的能力。

方法

使用钙荧光法评估农药对人精子细胞钙信号的影响。使用特异性 CatSper 抑制剂 RU1968 评估 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精子功能和活力的影响。

结果

在 10µM 时，发现 53 种农药中的 28 种能够诱导人精子细胞产生钙信号。这 28 种活性农药中的大多数通过 CatSper 诱导钙信号，并干扰随后由两种内源性 CatSper 配体孕酮和前列腺素 E 诱导的钙信号。发现多种活性农药在 10µM 时影响钙介导的精子功能和活力。单独或与其他环境化学品联合使用的活性农药的低 nM 剂量混合物被发现能够显著诱导钙信号，并抑制随后由孕酮和前列腺素 E 诱导的钙信号。

结论

我们的研究结果表明，农药无论是单独存在还是处于低 nM 剂量混合物中，都能在体外以低 nM 浓度干扰人精子细胞中的正常钙信号。在相关基质（如血液和生殖液）中对活性农药进行生物监测非常有限，因此，实时人类农药暴露对人精子细胞和生育力的影响在很大程度上仍然未知。人类农药暴露在多大程度上影响体内人类受精的成功机会需要进一步研究。