环境相关浓度磷酸三乙酯暴露对斑马鱼肝脏的毒性及其分子机制

doi:10.3969/j.issn.1004-616x.2026.02.007

摘要/Abstract

摘要： 目的： 探讨有机磷阻燃剂磷酸三乙酯(TEP)暴露对斑马鱼肝脏的毒性效应及机制，为TEP的生态风险评估提供科学依据。方法： 实验以4月龄AB系野生斑马鱼为实验动物，连续28 d暴露于环境相关浓度(0、1、10、100、1 000μg/L)的TEP中。暴露结束后，测定斑马鱼体重及体长，通过HE染色法观察肝脏组织结构的损伤，通过RT-qPCR、ELISA等方法检测肝脏功能指标(ALT和AST酶活性)、氧化应激和炎症相关指标(基因水平和酶活性)、代谢酶(CYP450)以及Keap1-Nrf2-ARE信号通路关键分子表达水平。结果： 与对照组相比，TEP暴露对斑马鱼体重及体长无明显影响。TEP暴露组斑马鱼肝脏HE病理切片显示肝血窦扩张与水肿，1、10μg/L TEP组见肝细胞核溶解，100、1 000μg/L TEP组同时见核溶解、核固缩及无核区；ALT、AST酶活性显著升高(P<0.05)。氧化应激相关基因Sod1、Sod2、Gstp1.1、Gstp1.2、Cat、Cyp1a、Cyp3a、Gpx1a的m RNA表达水平整体下调(P<0.05)，促炎因子基因IL-1β、TNF-α、IL-6的mRNA表达水平上调(P<0.05)。超氧化物歧化酶SOD、过氧化氢酶CAT、谷胱甘肽过氧化物酶GSH-Px、CYP450活性显著降低，丙二醛MDA含量显著升高(P<0.05)。Keap-Nrf2-ARE信号通路关键分子Keap1基因表达上调(P<0.05),Nrf2、Ho1、NQO1基因表达下调(P<0.05),Keap1、Nrf2、NQO1蛋白表达显著下调(P<0.05)。结论： 环境相关浓度TEP可能通过干扰Keap1-Nrf2-ARE信号通路诱导斑马鱼肝脏氧化应激损伤并伴随炎症反应，本文研究结果可为水体TEP污染的生态风险防控及相关毒理学研究提供科学参考。

关键词: 磷酸三乙酯, 斑马鱼, 环境相关浓度, 肝脏毒性, 分子机制

Abstract: OBJECTIVE: This study aimed to investigate liver toxicity of the organophosphate flame retardant triethyl phosphate(TEP) on zebrafish(Danio rerio), and to provide a scientific basis for the ecological risk assessment of TEP. METHODS: Four-month-old AB strain wild-type zebrafish were exposed to environmentally relevant concentrations of TEP(0, 1, 10, 100, and 1 000 μg/L) for 28 days. After exposure,body weight and body length were measured. Histopathological damage of the liver was observed via hematoxylin-eosin(HE) staining. Liver function indicators(activities of ALT and AST), oxidative stress and inflammation-related indicators(gene expression levels and enzyme activities),metabolic enzyme(CYP450),and expression levels of key molecules in the Keap1-Nrf2-ARE signaling pathway were detected using RT-qPCR,ELISA,and other methods. RESULTS: TEP exposure had no significant effect on the body weight or body length of zebrafish. HE pathological sections of exposed groups showed sinusoidal dilation and edema in the liver;nuclear lysis was observed in the 1 and 10 μg/L TEP groups,while nuclear lysis,nuclear pyknosis,and anuclear areas were simultaneously observed in the 100 and 1 000 μg/L groups. Activities of ALT and AST were significantly elevated. The mRNA expression of oxidative stress-related genes(Sod1,Sod2,Gstp1.1,Gstp1.2, Cat, Cyp1a, Cyp3a, and Gpx1a) were downregulated, while the mRNA expression of proinflammatory factors(IL-1β,TNF-α and IL-6) was upregulated. The activities of SOD,CAT,GSH-Px,and CYP450 were significantly decreased,whereas MDA content was significantly increased. For key molecules in the Keap1-Nrf2-ARE signaling pathway:the mRNA expression of Keap1 was upregulated,while Nrf2,Ho1,and NQO1 expressions were downregulated. Meanwhile, the protein expressions of Keap1, Nrf2 and NQO1 were significantly downregulated. CONCLUSION: Environmentally relevant concentrations of TEP induced oxidative stress injury accompanied by inflammatory response in zebrafish liver by interfering with the Keap1-Nrf2-ARE signaling pathway. The results provide scientific reference for ecological prevention and control of TEP pollution in water bodies.

Key words: triethyl phosphate, zebrafish, environmentally relevant concentrations, hepatotoxicity, molecular mechanism

中图分类号:

R994.6

吴平凡, 马雪, 曾晓琪, 朱强, 黄奇彦, 李红梅, 徐海明. 环境相关浓度磷酸三乙酯暴露对斑马鱼肝脏的毒性及其分子机制[J]. 癌变·畸变·突变, 2026, 38(2): 128-136.

WU Pingfan, MA Xue, ZENG Xiaoqi, ZHU Qiang, HUANG Qiyan, LI Hongmei, XU Haiming. Environmentally relevant concentrations of triethyl phosphate exposure on liver toxicity in zebrafish and its molecular mechanism[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2026, 38(2): 128-136.

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