马令怡, 李维溪, 尹洁晨, 朱光灿, 刘冉. 环境相关浓度的睾酮对秀丽线虫的生殖毒性作用[J]. 环境与职业医学, 2021, 38(8): 910-914. DOI: 10.13213/j.cnki.jeom.2021.20581
引用本文: 马令怡, 李维溪, 尹洁晨, 朱光灿, 刘冉. 环境相关浓度的睾酮对秀丽线虫的生殖毒性作用[J]. 环境与职业医学, 2021, 38(8): 910-914. DOI: 10.13213/j.cnki.jeom.2021.20581
MA Lingyi, LI Weixi, YIN Jiechen, ZHU Guangcan, LIU Ran. Reproductive toxicity of environmentally relevant concentrations of testosterone on Caenorhabditis elegans[J]. Journal of Environmental and Occupational Medicine, 2021, 38(8): 910-914. DOI: 10.13213/j.cnki.jeom.2021.20581
Citation: MA Lingyi, LI Weixi, YIN Jiechen, ZHU Guangcan, LIU Ran. Reproductive toxicity of environmentally relevant concentrations of testosterone on Caenorhabditis elegans[J]. Journal of Environmental and Occupational Medicine, 2021, 38(8): 910-914. DOI: 10.13213/j.cnki.jeom.2021.20581

环境相关浓度的睾酮对秀丽线虫的生殖毒性作用

Reproductive toxicity of environmentally relevant concentrations of testosterone on Caenorhabditis elegans

  • 摘要: 背景

    睾酮是一种重要的雄激素,由于人类生产生活活动造成它在环境中的不同程度暴露,可能影响生态环境中多种生物的生殖能力。

    目的

    以秀丽隐杆线虫为模式生物,探讨环境相关浓度的睾酮的生殖毒性作用。

    方法

    根据睾酮在环境中实际暴露水平选取有代表性的环境相关质量浓度(简称为浓度,1 ng·L-1、10 ng·L-1、100 ng·L-1、1 μg·L-1、10 μg·L-1)作为本研究的暴露浓度,将同步化的L1期野生型N2线虫暴露于不同浓度的睾酮及M9溶剂(对照组)中,20℃下恒温培养染毒48 h。观察各组生殖毒性指标,包括后代数目、世代时间、性腺发育阶段、畸形现象、生殖细胞数目。

    结果

    野生型N2秀丽隐杆线虫自L1期暴露于睾酮48 h后,与对照组(219.70±27.99)相比,10 ng·L-1~10 μg·L-1组后代数目减少(分别为186.90±19.68,P=0.009;182.60±25.33,P=0.003;178.20±14.46,P=0.001;159.20±28.64,P < 0.001)(F=7.24,P < 0.001);10 μg·L-1组与对照组相比后代数目减少了27.5%。与对照组(71.85±3.49)h相比,100 ng·L-1组(81.70±6.26)h,P=0.001、1 μg·L-1组(79.28±4.83)h,P=0.015、10 μg·L-1组(79.54±3.92)h,P=0.011世代时间延长(F=3.97,P=0.005);100 ng·L-1组世代时间延长最多,为9.85 h。对性腺发育阶段的观察发现,随染毒浓度的增高,性腺发育逐渐受到抑制,各个剂量组中的成虫比例从对照组的(63.82±16.37)%逐渐下降至(53.95±1.03)%、(41.39±8.57)%、(26.79±1.31)%、(20.16±1.94)%、(8.00±2.03)%。在100 ng·L-1及以上染毒组线虫中观察到畸形现象,包括排卵孔畸形、子宫内卵未受精、虫袋、性腺发育异常。与对照组(249.40±23.09)相比,10 ng·L-1~10 μg·L-1组生殖细胞数目减少分别为(224.50±28.48,P=0.048)、(215.20±27.28,P=0.005)、(194.30±26.78,P < 0.001)、(181.80±19.45,P < 0.001)(F=10.64,P < 0.001);10 μg·L-1组生殖细胞数目较对照组减少了27.1%。

    结论

    环境相关浓度的雄激素睾酮可以导致秀丽线虫后代数目减少、世代时间延长、性腺发育迟缓,这些作用可能与性腺中生殖细胞的减少有关。

     

    Abstract: Background

    Testosterone is an important androgen. However, human production and living activities have led to its exposure to varying degrees in the environment, which may affect the reproductive ability of many organisms in the ecological environment.

    Objective

    Caenorhabditis elegans (C. elegans) is used as a model organism to investigate the reproductive toxicity of testosterone at environmentally relevant concentrations.

    Methods

    According to the actual concentration of testosterone in the environment, synchronized L1-stage wild-type N2 nematodes were exposed to representative environmentally relevant concentrations (1 ng·L-1, 10 ng·L-1, 100 ng·L-1, 1 μg·L-1, 10 μg·L-1) of testosterone and M9 solvent (control) at 20℃ for 48 h. Brood size, generation time, gonadal development stage, malformation, and the number of germ cells were observed among the groups.

    Results

    After the wild-type N2 C. elegans being exposed to testosterone for 48 h in the L1 stage, compared with the control group (219.70±27.99), the brood sizes of C. elegans were significantly decreased in the 10 ng·L-1 to 10 μg·L-1 groups (186.90±19.68, P=0.009; 182.60±25.33, P=0.003; 178.20±14.46, P=0.001; 159.20±28.64, P < 0.001, respectively) (F=7.24, P < 0.001), and in the 10 μg·L-1 group the brood size decreased by 27.5%. Compared with the control group(71.85±3.49) h, the generation time was significantly prolonged in the 100 ng·L-1 group(81.70±6.26) h, P=0.001, 1 μg·L-1 group(79.28±4.83) h, P=0.015, and 10 μg·L-1 group(79.54±3.92) h, P=0.011 (F=3.97, P=0.005), and was longest in the 100 ng·L-1 group where the generation time was prolonged by 9.85 h. In terms of gonadal development stage, as the concentration of testosterone increased, the development of gonads was gradually inhibited, and the proportion of adult worms to total nematodes gradually decreased from (63.82±16.37)% in the control group to (53.95±1.03)%, (41.39±8.57)%, (26.79±1.31)%, (20.16±1.94)%, and (8.00±2.03)% in each dose group, respectively. Malformations were observed in the 100 ng·L-1 group and above, including ovulation hole malformation, unfertilized eggs in uterus, a bag of worm, and abnormal gonadal development. Compared with the control group (249.40±23.09), the numbers of germ cells were significantly declined in the 10 ng·L-1 to 10 μg·L-1 groups (224.50±28.48, P=0.048; 215.20±27.28, P=0.005; 194.30±26.78, P < 0.001; 181.80±19.45, P < 0.001, respectively) (F=10.64, P < 0.001), and in the 10 μg·L-1 group the number of germ cells decreased by 27.1%.

    Conclusion

    Androgen testosterone at environmentally relevant concentrations can lead to decreased brood size, prolonged generation time, and delayed gonadal development of C. elegans. These effects may be related to the decrease of germ cells.

     

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