Research on the endocrine disruption effect of typical phenolic pollutants: The embryonic development effects and molecule effects of gene regulation mediated by nuclear receptor on Zebrafish

SONG Jing-wen; JIN Ya-ru; LIU Hong-ling

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China Environmental Science ›› 2020, Vol. 40 ›› Issue (9) : 4065-4076.

Research on the endocrine disruption effect of typical phenolic pollutants: The embryonic development effects and molecule effects of gene regulation mediated by nuclear receptor on Zebrafish

SONG Jing-wen, JIN Ya-ru, LIU Hong-ling

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Abstract

2,4,6-trichlorophenol, 4-chloro-3-methylphenol and 4-nitrophenol were the three most abundant phenols in various environmental media around chemical industry park in Nanjing, China. In this study, zebrafish embryos were exposedto 0.1, 0.5, 2.5, 5 and 10mg/L of three phenols to observe the effects of morphological indicators in the early stages of embryonic development, and then we investigated the transcriptional profiles on 61 genes associated with 8 important receptors (androgen receptor (AR), thyroid receptor (TR), aryl hydrocarbon receptor (AhR), estrogen receptor (ER), glucocorticoid receptor (GR), pregnane Xreceptor (PXR), mineralocorticoid receptor (MR), peroxisome proliferator activated receptor (PPAR)) in zebrafish larvae by q-RT-PCR at 120 hpf. Moreover, the possible interaction processes between three phenols and their significantly affected nuclear receptors (including ER, AhR, PXR, GR and MR) were studied by in silico approaches (molecular docking and molecular dynamics simulation). Our results indicated that the three phenols at 5mg/L and 10mg/L concentrations all resulted in adverse effects on spinal curvature and edema in zebrafish embryos and larvae. The gene results of 2,4,6-trichlorophenol exposure could down-regulate gene of ER, the estrogen receptor α (er1) were down-regulated by 2.1-fold after exposure to 2.5mg/L of 2,4,6-trichlorophenol. 4-chloro-3-methylphenol significantly affected the genes of ER, AhR and PXR. er1 was significantly down-regulated by 1.8-fold and 2.2-fold respectively, at 0.5 and 2.5mg/L, and aryl hydrocarbon receptor gene 2 (ahr2) was down-regulated by 1.7-fold and 1.8-fold, and pregnane X receptor gene (pxr) was up-regulated by 3.5-fold and 3.2-fold at 0.1 and 2.5mg/L.Exposure to 4-nitrophenol caused regulatory changes in the core receptor genes of GR and MR. The glucocorticoid receptor gene (gr) was significantly up-regulated by 2.0-fold and 2.1-fold at concentrations of 0.1 and 0.5mg/L, and down-regulated by 1.9-fold at 2.5mg/L. The glucocorticoid receptor gene (mr) was down-regulated by 1.9-fold and 2.1-fold at concentrations of 0.5 and 2.5mg/L, and significantly up-regulated by 1.7-fold under the exposure concentration of 0.1mg/L.In silico results showed that the three phenols were able to dock into the ligand-binding domain and stably bound to the related receptors through hydrophobic and hydrogen bonding interactions. Also, relative root mean square deviation (RMSD) fluctuations for the backbone atoms was stable after 5ns. This study concluded that these three phenols could mediate endocrine disrupting effects and affect the early growth and development of zebrafish through ER, AhR, PXR, GR and MR receptors.

Key words

2,4,6-trichlorophenol / 4-chloro-3-methylphenol / 4-nitrophenol / embryonic development toxicity / molecular dynamics simulation / nuclear receptor

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SONG Jing-wen, JIN Ya-ru, LIU Hong-ling. Research on the endocrine disruption effect of typical phenolic pollutants: The embryonic development effects and molecule effects of gene regulation mediated by nuclear receptor on Zebrafish[J]. China Environmental Science. 2020, 40(9): 4065-4076

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