宋静文,
靳亚茹,
刘红玲
南京大学环境学院, 污染控制与资源化研究国家重点实验室, 南京 210023

作者简介: 宋静文(1995-),男,硕士研究生,研究方向为环境毒理学,E-mail:jingwensong@yeah.net.

基金项目: 国家自然科学基金资助项目(21677073)；国家重点研发项目(2018YFC1801505)；国家科技重大专项(2017ZX07301002，2018ZX07208001)


中图分类号: X171.5

Endocrine Disruption Effect of BDE-28 Mediated by Zebrafish Nuclear Receptor

Song Jingwen,
Jin Yaru,
Liu Hongling
State Key Laboratory of Pollution Control & Resource Reuse, School of the Environment, Nanjing University, Nanjing 210023, China

CLC number: X171.5

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摘要:2,4,4’-三溴联苯醚(BDE-28)在环境中普遍存在，且在长江流域多种水生生物中检出。目前，国内外对高溴代PBDEs(如BDE-47、BDE-99等)的水生脊椎动物内分泌干扰效应报道较多，而BDE-28的有关研究则相对较少。将斑马鱼胚胎暴露于2、20和200 μg·L^-1的BDE-28后，借助q-RT-PCR方法对幼鱼8个重要受体包括雄激素受体(AR)、甲状腺激素受体(TR)、芳香烃受体(AhR)、雌激素受体(ER)、糖皮质激素受体(GR)、孕烷X受体(PXR)、盐皮质激素受体(MR)和过氧化物酶体增殖物激活受体(PPAR)相关基因的转录水平进行了研究。结果表明，BDE-28暴露可导致AR、TR和AhR的基因下调，其中核心受体AR和TR在低中高3种浓度下的下调倍数分别为3.03、2.64、10.10和2.21、2.18、2.31，芳香烃受体基因2(ahr2)在暴露于2和20 μg·L^-1的BDE-28后，下调倍数分别为12.65和9.23，而雌激素受体(er1)基因在低中高浓度显著上调，上调倍数分别为12.29、12.67和15.87，雌激素受体(er2a)基因在2和20 μg·L^-1 BDE-28下的上调倍数分别为10.83和17.19。进一步采用分子对接和分子动力学模拟的方法研究BDE-28与AR、TR、AhR和ER之间的相互作用。结果显示，BDE-28与这些受体通过疏水和氢键等相互作用稳定结合，动力学模拟后骨架原子的均方根偏差(RMSD)在5 ns后较稳定。由此可知，BDE-28通过AR、TR、AhR和ER受体介导产生内分泌干扰效应。
关键词: BDE-28/
斑马鱼/
内分泌干扰/
核受体/
分子对接/
分子动力学模拟

Abstract:2,4,4’-tribromodiphenyl ether (BDE-28) is ubiquitous in the environment and has been found in a variety of aquatic organisms in Yangtze River basin. Compared with high brominated PBDEs (such as BDE-47, BDE-99), research on endocrine disrupting effect of BDE-28 was few in aquatic vertebrates. In our study, zebrafish embryos were exposed to 2, 20 and 200 μg·L^-1 BDE-28, and the transcriptional profiles of 41 genes associated with 8 important receptors were investigated with a q-RT-PCR assay, including androgen receptor (AR), thyroid hormone receptor (ThR), aryl hydrocarbon receptor (AhR), estrogen receptor (ER), glucocorticoid receptor (GR), pregnane X receptor (PXR), mineralocorticoid receptor (MR) and peroxisome proliferator activated receptor (PPAR). Genes related to AR, ThR and AhR were significantly down-regulated while genes related to ER were significantly up-regulated. Genes of AR and TR were down-regulated by 3.03-, 2.64-, 10.10- and 2.21-, 2.18-, 2.31-fold after exposure to 2, 20 and 200 μg·L^-1 BDE-28, respectively, while the aryl hydrocarbon receptor 2 gene (ahr2) was down-regulated by 12.65-fold at 2 μg·L^-1 BDE-28 and 9.23-fold at 20 μg·L^-1 BDE-28. Estrogen receptor (er1) gene was significantly up-regulated by 12.29-, 12.67- and 15.87-fold, respectively, at 2, 20 and 200 μg·L^-1 BDE-28, and estrogen receptor (er2a) gene was induced by 10.83- and 17.19-fold, respectively, at 2 and 20 μg·L^-1 BDE-28. The possible interactions between BDE-28 and these four receptors were further studied by using molecular docking and molecular dynamics simulation. The simulation showed that BDE-28 bound stably to these receptors through hydrophobic interactions and hydrogen bonds. Also, the relative root means square deviation (RMSD) of the backbone atoms became steady after 5 ns in the simulation. It can be concluded that BDE-28 causes endocrine disruption through AR, TR, AhR and ER-mediated pathways.
Key words:BDE-28/
zebrafish/
endocrine disruption/
nuclear receptor/
molecular docking/
molecular dynamics simulation.

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