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孕期邻苯二甲酸酯暴露对小鼠精子获能参数的影响。

Effects of preconception exposure to phthalates on mouse sperm capacitation parameters.

机构信息

Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, Massachusetts, USA.

C.S. Mott Center for Human Growth and Development, Department of Obstetrics and Gynecology, School of Medicine, Wayne State University, Detroit, Michigan, USA.

出版信息

Andrology. 2023 Oct;11(7):1484-1494. doi: 10.1111/andr.13423. Epub 2023 Mar 21.

Abstract

BACKGROUND

Phthalates have been linked to adverse male reproductive health, including poor sperm quality and embryo quality as well as a longer time to pregnancy (months of unprotected intercourse before conception occurs). The present study aimed to evaluate the effect of preconception exposure to two ubiquitous phthalate chemicals, di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP), and their mixture on sperm function, fertilization, and embryo development in mice.

MATERIALS AND METHODS

Adult male C57BL/6J mice aged 8-9 weeks were exposed to di(2-ethylhexyl) phthalate, di-n-butyl phthalate, or their mixture (di-n-butyl phthalate + di(2-ethylhexyl) phthalate) at 2.5 mg/kg/day or vehicle for 40 days (equivalent to one spermatogenic cycle) via surgically implanted osmotic pumps. Caudal epididymal spermatozoa were extracted and analyzed for motility using computer-assisted sperm analyses. Sperm phosphorylation of protein kinase A substrates and tyrosine phosphorylation, markers of early and late capacitation events, respectively, were analyzed by Western blots. In vitro fertilization was used to evaluate the sperm fertilizing capacity.

RESULTS

While the study did not reveal any significant differences in sperm motility and fertilization potential, abnormal sperm morphology was observed in all phthalate exposures, particularly in the phthalate mixture group. In addition, the study revealed significant differences in sperm concentration between control and exposed groups. Moreover, protein phosphorylation of protein kinase A substrates was decreased in the di(2-ethylhexyl) phthalate and mixture exposure groups, while no significant changes in protein tyrosine phosphorylation were observed in any of the groups. Assessment of the reproductive functionality did not reveal significant effects on in vitro fertilization and early embryo development rates but showed wide variability in the phthalate mixture group.

CONCLUSION

Our findings suggest that preconception phthalate exposure affects sperm numbers and phosphorylation of protein kinase A substrates involved in capacitation. Future research is warranted to examine the associations between phthalate exposure and capacitation in human spermatozoa.

摘要

背景

邻苯二甲酸酯已被证实与男性生殖健康不良有关,包括精子质量和胚胎质量下降,以及妊娠时间延长(即从无保护性行为开始到受孕发生需要数月时间)。本研究旨在评估孕前暴露于两种常见邻苯二甲酸酯化学物质,即邻苯二甲酸二(2-乙基己基)酯(DEHP)、邻苯二甲酸二正丁酯(DBP)及其混合物对小鼠精子功能、受精和胚胎发育的影响。

材料和方法

8-9 周龄成年雄性 C57BL/6J 小鼠通过手术植入的渗透泵以 2.5mg/kg/天或载体的剂量分别暴露于邻苯二甲酸二(2-乙基己基)酯、邻苯二甲酸二正丁酯或其混合物(邻苯二甲酸二正丁酯+邻苯二甲酸二(2-乙基己基)酯)40 天(相当于一个精子发生周期)。提取尾附睾精子并使用计算机辅助精子分析进行运动分析。通过 Western blot 分析蛋白激酶 A 底物的精子磷酸化和酪氨酸磷酸化,分别作为早期和晚期获能事件的标志物。体外受精用于评估精子的受精能力。

结果

尽管该研究未发现精子运动和受精能力有任何显著差异,但在所有邻苯二甲酸酯暴露组中均观察到精子形态异常,尤其是在邻苯二甲酸酯混合物组中。此外,研究还发现对照组和暴露组之间的精子浓度存在显著差异。此外,蛋白激酶 A 底物的蛋白磷酸化在邻苯二甲酸二(2-乙基己基)酯和混合物暴露组中减少,而在任何组中均未观察到蛋白酪氨酸磷酸化的显著变化。对生殖功能的评估并未显示对体外受精和早期胚胎发育率有显著影响,但在邻苯二甲酸酯混合物组中显示出广泛的变异性。

结论

我们的研究结果表明,孕前邻苯二甲酸酯暴露会影响精子数量和参与获能的蛋白激酶 A 底物的磷酸化。需要进一步的研究来检测人类精子中邻苯二甲酸酯暴露与获能之间的关联。

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