2-乙基己基二苯基磷酸酯急性暴露对斑马鱼仔鱼脂质代谢和线粒体功能的影响
Effects of Acute Exposure to 2-ethylhexyl Diphenyl Phosphate on Lipid Metabolism and Mitochondrial Function in Zebrafish Larvae
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摘要: 2-乙基己基二苯基磷酸酯(2-ethylhexyl diphenyl phosphate, EHDPP)是一种典型的有机磷阻燃剂，该化合物的大量使用导致其在环境基质中被频繁检出。大量离体研究表明EHDPP具有干扰脂质代谢的不良影响，但其是否会引起活体脂质代谢紊乱及潜在的机制尚不明确。本研究将斑马鱼胚胎(2 hours post-fertilization, 2 hpf)暴露在环境相关浓度(0、1、10、100、250和500 μg·L-1)EHDPP中5 d，评估EHDPP对仔鱼脂质代谢和线粒体功能的影响。结果表明，和对照组相比，100、250和500 μg·L-1 EHDPP暴露显著降低仔鱼孵化率、体长、心率和存活率，畸形率升高，表现出显著的生长发育毒性。结果还显示EHDPP引发脂质紊乱，总胆固醇和甘油三酯的含量在500 μg·L-1 EHDPP暴露组较对照组显著性降低。EHDPP暴露显著降低仔鱼三磷酸腺苷(adenosine triphosphate, ATP)、超氧化物歧化酶(superoxide dismutase, SOD)和谷胱甘肽(glutathione, GSH)水平，活性氧(reactive oxygen species, ROS)水平显著性升高，这提示EHDPP可能会诱发线粒体功能障碍。进一步发现EHDPP暴露导致仔鱼18rs-rrn基因表达显著性下降，而参与调控线粒体分裂(线粒体动力相关蛋白(drp1)和线粒体分裂蛋白(fis1))、融合(视神经萎缩蛋白(opa1))和自噬(PTEN诱导激酶1(pink1)和FUN14域蛋白1(fundc1))的基因表达显著性升高，表明EHDPP引发线粒体功能障碍。参与脂质合成的基因(脂肪酸合酶基因(fas)和载脂蛋白E基因(apoe))显著性降低，而调控脂质代谢的基因，如过氧化物酶体增殖物激活受体α基因(pparα)、过氧化物酶体增殖物激活受体β基因(pparβ)和酰基辅酶A氧化酶1基因(acox1)显著性升高，这些结果表明EHDPP可能引发斑马鱼线粒体功能障碍进而干扰斑马鱼幼鱼的脂肪酸氧化和脂质合成，总的来说，本研究发现EHDPP可能诱发脂质紊乱，并首次在体内实验中表明EHDPP会引发斑马鱼线粒体功能障碍，为EHDPP的毒性机制的研究提供新的思路。
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关键词:
- 2-乙基己基二苯基磷酸酯 /
- 斑马鱼 /
- 脂质紊乱 /
- 线粒体功能障碍
Abstract: 2-ethylhexyl diphenyl phosphate (EHDPP) is an aryl organophosphorus flame retardant, which is ubiquitously found in various environmental matrices. While previous in vitro studies have emphasized the negative impact of EHDPP on lipid metabolism, there remains a dearth of in vivo research exploring its potential toxic effects and underlying mechanisms. In the present study, zebrafish (Danio rerio) embryos (<2 hpf) were exposed to environmentally relevant concentrations (0, 1, 10, 100, 250 and 500 μg·L-1) for 5 days. Results showed that 100, 250 and 500 μg·L-1 EHDPP may lead to developmental toxicity, evidenced by the increased malformation rate, reduced hatching rate, heart rate, survival rate and body length, in comparison with the control. The levels of total cholesterol and triglycerides were significantly reduced in 500 μg·L-1 EHDPP group compared to the control, suggesting that EHDPP caused disorders of lipid metabolism. EHDPP resulted in a notable reduction in the concentrations of ATP, SOD, and GSH, along with an elevation in ROS levels, suggestive of mitochondrial dysfunction triggered by EHDPP. Upon further analysis, it was revealed that the transcription of the 18rs-rrn gene was significantly reduced, while the expressions of genes involved in mitochondrial division (including mitochondrial fission protein 1 (fis1) and dynamin-related protein (drp1)), fusion (optic atrophy 1 (opa1)), and autophagy (PTEN-induced putative kinase 1 (pink1) and FUN14 domain containing 1 (fundc1)) were notably upregulated, which may partially account for the mitochondrial dysfunction. Genes involved in lipid biosynthesis including acid synthetase (fas) and apolipoprotein E (apoe) genes were significantly downregulated, whereas genes related to lipid metabolism such as peroxisome proliferators-activated receptors (pparα and pparβ) and acyl-CoA oxidase 1 (acox1) were upregulated. These results implied that EHDPP may disturb fatty acid oxidation and lipid synthesis in zebrafish larvae which might result from mitochondrial dysfunction. In conclusion, our study found that EHDPP may induce lipid metabolism disorders. For the first time, we have demonstrated mitochondrial dysfunction in vivo resulting from the effects of EHDPP, potentially offering novel insights into the toxicity elicited by this compound. -
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