Hong Taeyeon, Park Hahyun, An Garam, Song Gwonhwa, Lim Whasun
Department of Biological Sciences, College of Science, Sungkyunkwan University, Suwon 16419, Republic of Korea.
Institute of Animal Molecular Biotechnology and Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Republic of Korea.
Sci Total Environ. 2023 Jan 1;854:158780. doi: 10.1016/j.scitotenv.2022.158780. Epub 2022 Sep 14.
Ethalfluralin, of dinitroaniline herbicide family, is an effective weed controller. Following residue detection in herbicide-treated fields, ethalfluralin was reported to interfere with early stages of implantation in some vertebrate species. However, the role of ethalfluralin in the development of zebrafish embryos has not been elucidated yet. Therefore, in the present study, we investigated the morphological and physiological changes that occur in the embryonic development of zebrafish due to ethalfluralin exposure. Results indicated that ethalfluralin decreased survival rate along with reduction in the hatching ratio and heartbeat. It was observed to cause edema in the heart and yolk sac, and apoptosis in the anterior region of the developing zebrafish larvae; as visualized through acridine orange and TUNEL staining. In addition, ethalfluralin increased the expression of the apoptosis-associated genes including tp53, cyc1, casp8, casp9, and casp3. The Seahorse Mito Stress analysis revealed that ethalfluralin slightly reduced mitochondrial respiration in live zebrafish embryos. Reactive oxygen species (ROS) production was also observed to be elevated in zebrafish larvae in response to ethalfluralin. Treatment with ethalfluralin decreased blood vessel formation in brain and intestine in flk1 transgenic zebrafish embryos. The decrease in angiogenesis related gene expression was specifically observed in vegfc, flt1, and kdrl, and in the intestinal vasculature related genes apoa4a, aqp3, fabp2, and vil1. Moreover, an increase in inflammatory genes such as cox2a, cox2b, cxcl-c1c, il8, mcl1a, mcl1b, and nf-κb was observed using real-time PCR analysis. Collectively, these results indicate that oxidative stress generated by exposure to ethalfluralin induced ROS generation, apoptosis, inflammation and anti-angiogenic effects, and therefore, ethalfluralin may be toxic to the development of zebrafish embryos.
乙丁氟灵属于二硝基苯胺类除草剂，是一种有效的杂草控制剂。在除草剂处理过的田地中检测到残留后，有报道称乙丁氟灵会干扰某些脊椎动物物种的早期着床阶段。然而，乙丁氟灵在斑马鱼胚胎发育中的作用尚未阐明。因此，在本研究中，我们调查了由于暴露于乙丁氟灵而在斑马鱼胚胎发育过程中发生的形态和生理变化。结果表明，乙丁氟灵降低了存活率，同时孵化率和心跳也降低。观察到它会导致心脏和卵黄囊水肿，并在发育中的斑马鱼幼虫前部区域诱导凋亡；通过吖啶橙和TUNEL染色可以观察到。此外，乙丁氟灵增加了包括tp53、cyc1、casp8、casp9和casp3在内的凋亡相关基因的表达。海马线粒体应激分析表明，乙丁氟灵会轻微降低活斑马鱼胚胎中的线粒体呼吸。还观察到，响应乙丁氟灵，斑马鱼幼虫中的活性氧（ROS）产生增加。用乙丁氟灵处理会减少flk1转基因斑马鱼胚胎中脑和肠道的血管形成。在vegfc、flt1和kdrl以及肠道血管相关基因apoa4a、aqp3、fabp2和vil1中特别观察到血管生成相关基因表达的下降。此外，使用实时PCR分析观察到炎症基因如cox2a、cox2b、cxcl-c1c、il8、mcl1a、mcl1b和nf-κb的表达增加。总的来说，这些结果表明，暴露于乙丁氟灵产生的氧化应激会诱导ROS产生、凋亡、炎症和抗血管生成作用，因此，乙丁氟灵可能对斑马鱼胚胎的发育有毒性。
