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膳食暴露于 17α-乙炔基雌二醇是否会改变成年三倍体斑马鱼中与内分泌干扰和氧化应激相关的生物标志物?

Does dietary exposure to 17α-ethinylestradiol alter biomarkers related with endocrine disruption and oxidative stress in the adult triploid of Danio rerio?

机构信息

Department of Animal Protection and Welfare and Veterinary Public Health, University of Veterinary Sciences Brno, Palackeho tr. 1946/1, 612 42 Brno, Czech Republic.

Department of Animal Protection and Welfare and Veterinary Public Health, University of Veterinary Sciences Brno, Palackeho tr. 1946/1, 612 42 Brno, Czech Republic.

出版信息

Sci Total Environ. 2023 Apr 20;870:161911. doi: 10.1016/j.scitotenv.2023.161911. Epub 2023 Jan 31.

Abstract

This study was conducted to investigate a comprehensive effect of 17α-ethinylestradiol (EE2) in zebrafish (Danio rerio) with the emphasis on endocrine disruption, oxidative stress and detoxification processes at different levels. Adult male triploid zebrafish were exposed to EE2 administered in feed at two concentrations - 10 and 1000 μg/kg for six weeks. The estrogenic potential of EE2 was evaluated using an analysis of vitellogenin, gene expression focused on reproductive disorders and gonad histological examination. The alterations in antioxidant and detoxification status were assessed using analyses of enzyme activities and changes in transcriptional levels of selected genes. The most significant changes were observed especially in fish exposed to a high concentration of EE2 (i.e., 1000 μg/kg). Such high concentration caused extensive mortality (25 %) mainly in the second half of the experiment followed by a highly significant decrease in the length and body weight. Similarly, highly significant induction of vitellogenin level and vtg1 mRNA expression (about 43,000-fold compared to the control) as well as a significant downregulation of gonad aromatase expression (cyp19a1a) and histological changes in testicular tissue were confirmed in this group. In the group exposed to environmentally relevant concentration of EE2 (i.e., 10 μg/kg), no significant differences in vitellogenin were observed, although all fish were positive in the detection of vitellogenin compared to control, where only 40 % of individuals were positive. In addition, the high concentration of EE2 resulted in significant alterations in most monitored antioxidant and detoxifying enzymes with the exception of catalase, followed by strongly significant upregulation in mRNA expression of gsr, gpx1a, cat and cyp1a genes. Furthermore, a significant decrease in the glutathione reductase activity was recorded in fish exposed to 10 μg EE2/kg. To our knowledge, this is the first study which reports the effects of subchronic per oral exposure to EE2 in adult triploid zebrafish.

摘要

本研究旨在探讨 17α-乙炔基雌二醇(EE2)对斑马鱼(Danio rerio)的综合影响,重点关注不同水平的内分泌干扰、氧化应激和解毒过程。将成年雄性三倍体斑马鱼用饲料暴露于 EE2 中,浓度分别为 10 和 1000μg/kg,持续 6 周。采用分析卵黄蛋白原、生殖障碍相关基因表达和性腺组织学检查来评估 EE2 的雌激素潜力。通过分析酶活性和选定基因转录水平的变化来评估抗氧化和解毒状态的改变。在暴露于高浓度 EE2(即 1000μg/kg)的鱼类中观察到最显著的变化。这种高浓度导致广泛的死亡率(25%),主要发生在实验的后半段,随后长度和体重显著下降。同样,在该组中,卵黄蛋白原水平和 vtg1 mRNA 表达显著诱导(与对照组相比约 43000 倍),以及性腺芳香化酶表达(cyp19a1a)和睾丸组织学变化显著下调,证实了这一点。在暴露于环境相关浓度 EE2(即 10μg/kg)的组中,未观察到卵黄蛋白原的显著差异,尽管与对照组相比,所有鱼类的卵黄蛋白原检测均呈阳性,而对照组中只有 40%的个体呈阳性。此外,除了过氧化氢酶外,高浓度 EE2 导致大多数监测到的抗氧化和解毒酶发生显著改变,随后 gsr、gpx1a、cat 和 cyp1a 基因的 mRNA 表达显著上调。此外,在暴露于 10μg EE2/kg 的鱼类中记录到谷胱甘肽还原酶活性显著下降。据我们所知,这是首次报道亚慢性口服暴露于成年三倍体斑马鱼的 EE2 的影响。

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