Dasgupta Tiasha, Manickam Venkatraman, Tamizhselvi Ramasamy
Department of Biosciences, School of Biosciences and Technology, Vellore Institute of Technology, Vellore, Tamil Nadu, India.
Sci Rep. 2025 Mar 17;15(1):9066. doi: 10.1038/s41598-025-93539-8.
Fetal alcohol spectrum disorders (FASD) are a group of physical, behavioral, and cognitive impairments caused by ethanol exposure during pregnancy. Zebrafish have emerged as a useful model for researching FASD and its variants in recent years. Oxidative stress has been identified as the primary damaging pathway, notwithstanding the possibility of other mechanisms at play. In this regard it's important to put an effort towards antioxidants which can exhibits and bare a potential to counteract the oxidative stress induced by ethanol during embryos development. Previosuly benzydamine has shown to protect macrophages against ethanol-induced condition by stabilizing redox homeostatis. This study aims to repurpose the Non-Steroidal Anti-Inflammatory Drug (NSAID) benzydamine to mitigate ethanol-induced teratogenesis during the early embryonic stage in Zebrafish. Zebrafish embryos were treated with 1% ethanol at 2 h post fertilization (hpf) and co-exposed with benzydamine (5-20 µM) after 2 h of ethanol treatment for 24 h. Reactive oxygen Species (ROS) and biochemical analysis was carried out at 48hpf. 1% ethanol significantly increased the production of ROS along with increased in lipid peroxidation followed by a decrease in glutathione (GSH) level when compoared to the control group (P < 0.001). These conditions were positively encountered by benzydamine (10, 15 µM) and returned to basal level. Involvement of two ethanol metabolizing enzymes cyp2y3 (Cytochrome P450, family 2, subfamily Y, polypeptide 3) and cyp3a65 Cytochrome P450, family 3, subfamily A, polypeptide 65 ) were also studied at 48hpf. 1% ethanol exposure aggregately elevated the expression of these two enzymes which showed a significant decrease in the benzydamine treated groups. Furthermore, the malformations and cellular damage due to 1% ethanol was studied at 96hpf, where 1% ethanol made severe malformation along with muscle fiber alteration, apoptosis in the brain and eye as manifested. These conditions were successfully reverted by benzydamine. In conclusion, ethanol causes oxidative stress, cellular damage along with severe malformation at early embryonic stage, which were partially prevented by the exposure of benzydamine.
胎儿酒精谱系障碍（FASD）是一组因孕期接触乙醇而导致的身体、行为和认知障碍。近年来，斑马鱼已成为研究FASD及其变体的有用模型。尽管可能还有其他机制在起作用，但氧化应激已被确定为主要的损伤途径。在这方面，努力寻找能够展现并具有抵消胚胎发育过程中乙醇诱导的氧化应激潜力的抗氧化剂非常重要。此前已表明，苄达明通过稳定氧化还原稳态来保护巨噬细胞免受乙醇诱导的损伤。本研究旨在重新利用非甾体抗炎药苄达明，以减轻斑马鱼早期胚胎阶段乙醇诱导的致畸作用。在受精后2小时（hpf）用1%乙醇处理斑马鱼胚胎，并在乙醇处理2小时后与苄达明（5 - 20 μM）共同暴露24小时。在48hpf进行活性氧（ROS）和生化分析。与对照组相比，1%乙醇显著增加了ROS的产生，同时脂质过氧化增加，随后谷胱甘肽（GSH）水平降低（P < 0.001）。苄达明（10、15 μM）积极应对了这些情况，使其恢复到基础水平。在48hpf还研究了两种乙醇代谢酶cyp2y3（细胞色素P450 2Y3）和cyp3a65（细胞色素P450 3A65）的参与情况。1%乙醇暴露总体上提高了这两种酶的表达，而在苄达明处理组中则显著降低。此外，在96hpf研究了1%乙醇导致的畸形和细胞损伤，1%乙醇导致了严重畸形以及肌肉纤维改变、脑和眼的细胞凋亡。这些情况被苄达明成功逆转。总之，乙醇在早期胚胎阶段会导致氧化应激、细胞损伤以及严重畸形，而苄达明的暴露可部分预防这些情况。
