Gu Kaicheng, Yang Lindong, Jiang Yi, Wang Zhiqiang, Chen Jiannan
School of life Science, Nanjing Normal University, No. 1 Wenyuan Road, Qixia District, Nanjing 210023, China.
Department of Obstetrics and Gynecology, School of Medicine, Nanjing University, 305 Zhongshan East Road, Xuanwu District, Nanjing 210018, China.
Biology (Basel). 2025 Aug 4;14(8):992. doi: 10.3390/biology14080992.
As typical environmental hormones, endocrine-disrupting chemicals (EDCs) have become a global environmental health issue of high concern due to their property of interfering with the endocrine systems of organisms. As a commonly used substitute for bisphenol A (BPA), bisphenol E (BPE) has been frequently detected in environmental matrices such as soil and water in recent years. Existing research has unveiled the developmental and reproductive toxicity of BPE; however, only one in vitro cellular experiment has preliminarily indicated potential neurotoxic risks, with its underlying mechanisms remaining largely unelucidated in the current literature. Potential toxic mechanisms and action targets of BPE were predicted using the zebrafish model via network toxicology and molecular docking, with RT-qPCRs being simultaneously applied to uncover neurotoxic effects and associated mechanisms of BPE. A significant decrease ( < 0.05) in the frequency of embryonic spontaneous movements was observed in zebrafish at exposure concentrations ≥ 0.01 mg/L. At 72 hpf and 144 hpf, the larval body length began to shorten significantly from 0.1 mg/L to 1 mg/L, respectively ( < 0.01), accompanied by a reduced neuronal fluorescence intensity and a shortened neural axon length ( < 0.01). By 144 hpf, the motor behavior in zebrafish larvae was inhibited. Through network toxicology and molecular docking, HSP90AB1 was identified as the core target, with the cGMP/PKG signaling pathway determined to be the primary route through which BPE induces neurotoxicity in zebrafish larvae. BPE induces neuronal apoptosis and disrupts neurodevelopment by inhibiting the cGMP/PKG signaling pathway, ultimately suppressing the larval motor behavior. To further validate the experimental outcomes, we measured the expression levels of genes associated with neurodevelopment (, , , ), serotonergic synaptic signaling (, ), the cGMP/PKG pathway (), and apoptosis (, ). These results offer crucial theoretical underpinnings for evaluating the ecological risks of BPE and developing environmental management plans, as well as crucial evidence for a thorough comprehension of the toxic effects and mechanisms of BPE on neurodevelopment in zebrafish larvae.
作为典型的环境激素，内分泌干扰化学物质（EDCs）因其具有干扰生物体内分泌系统的特性，已成为全球高度关注的环境健康问题。作为双酚A（BPA）的常用替代品，双酚E（BPE）近年来在土壤和水等环境基质中频繁被检测到。现有研究揭示了BPE的发育和生殖毒性；然而，仅有一项体外细胞实验初步表明其存在潜在神经毒性风险，目前文献中其潜在机制仍 largely未阐明。通过网络毒理学和分子对接，利用斑马鱼模型预测了BPE的潜在毒性机制和作用靶点，同时应用实时定量聚合酶链反应（RT-qPCR）来揭示BPE的神经毒性效应及相关机制。在暴露浓度≥0.01 mg/L时，斑马鱼胚胎自发运动频率显著降低（<0.05）。在72小时胚胎期（hpf）和144小时胚胎期，幼虫体长分别从0.1 mg/L和1 mg/L开始显著缩短（<0.01），同时伴有神经元荧光强度降低和神经轴突长度缩短（<0.01）。到144 hpf时，斑马鱼幼虫的运动行为受到抑制。通过网络毒理学和分子对接，确定热休克蛋白90αB1（HSP90AB1）为核心靶点，环磷酸鸟苷/蛋白激酶G（cGMP/PKG）信号通路为BPE诱导斑马鱼幼虫神经毒性的主要途径。BPE通过抑制cGMP/PKG信号通路诱导神经元凋亡并破坏神经发育，最终抑制幼虫运动行为。为进一步验证实验结果，我们检测了与神经发育相关的基因（，，，）、5-羟色胺能突触信号相关基因（，）、cGMP/PKG通路相关基因（）以及凋亡相关基因（，）的表达水平。这些结果为评估BPE的生态风险和制定环境管理计划提供了关键的理论基础，也为深入理解BPE对斑马鱼幼虫神经发育的毒性效应和机制提供了关键证据。
