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Sex-specific effects of fluoride and lead exposures on histology, antioxidant physiology, and immune system in the liver of zebrafish (Danio rerio)


Fluoride and Pb are both toxic to organisms; however, their combination effects and the corresponding toxic mechanisms remain unclear. In this study, male and female zebrafish (1:1) were evaluated to understand the effects of F and Pb alone and combined on growth, tissue microstructure, oxidative stress, and immune system functions of the liver. Four different groups and two exposure periods were compared: control group (C group), 80 mg/L fluoride group (F group), 60 mg/L lead group (Pb group), and 80 mg/L fluoride + 60 mg/L lead group (F + Pb group) for 45 and 90 days. The results indicated that F and Pb reduced growth performances; F + Pb treatment inhibited the growth performance traits of male zebrafish more than those of female zebrafish. Histopathological examination revealed large areas with focal necrosis, hepatocytes with karyolysis, and pycnotic nuclei in zebrafish exposed to F and Pb. The oxidative balance indices in the liver in the F and Pb groups were disturbed. F + Pb co-exposure aggravated oxidative stress in a time-dependent manner. The most serious oxidative stress was observed in the male zebrafish of the F + Pb group. Moreover, F and Pb exposure of male zebrafish increased pro-inflammatory and anti-inflammatory cytokines expression, which was decreased after 90 days of exposure. These results demonstrated that both F and Pb could damage the liver via downstream alterations in the activities of immune-related enzymes and in the levels of immune-related genes. F and Pb showed synergistic or additive effects. Male zebrafish were found to be more sensitive to F and Pb than female zebrafish.

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Data availability

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.


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This study was supported by the Shanxi Scholarship Council of China (2020-061), the Shanxi Provincial Key Research and Development Project (201903D221009), and the National Natural Science Foundation of China (31502141; 31440087).

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GW: data curation, writing—original draft preparation, methodology. TW: data curation, conceptualization, methodology. XZ: formal analysis, validation. JC: data curation, methodology; CF: conceptualization; SY: formal analysis; YC: formal analysis; FC: methodology; JC: supervision, project administration, writing—reviewing and editing.

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Correspondence to Jinling Cao.

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Wang, G., Wang, T., Zhang, X. et al. Sex-specific effects of fluoride and lead exposures on histology, antioxidant physiology, and immune system in the liver of zebrafish (Danio rerio). Ecotoxicology 31, 396–414 (2022).

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  • Fluoride
  • Lead
  • Liver
  • Sex-specific effect
  • Antioxidant physiology
  • Immune systems