Zhao Xiaoyu, Zhang Yuanyuan, Yu Ting, Cai Ling, Liang Junlang, Chen Zhong, Pan Chenyuan, Yang Ming
School of Environmental and Chemical Engineering, Shanghai University, Shanghai 200444, China.
Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, Fujian 361005, China.
Ecotoxicol Environ Saf. 2023 Aug 7;262:115324. doi: 10.1016/j.ecoenv.2023.115324.
Bisphenol AF (BPAF) is an emerging endocrine-disrupting chemical (EDC) prevalent in the environment as one of the main substitutes for bisphenol A. Sex-specific effects of EDCs have been commonly reported and closely linked to sexually dimorphic patterns of hormone metabolism and related gene expression during different exposure windows, but our understanding of these mechanisms is still limited. Here, following 28-day exposure of adult zebrafish to an environmentally relevant concentration of BPAF at 10 μg/L, the global transcriptional networks applying RNA sequencing (RNA-seq) and Ingenuity Pathway Analysis (IPA) were respectively investigated in the male and female fish liver, connecting the sex-dependent toxicity of the long-term exposure of BPAF to molecular responses. As a result, more differentially expressed genes (DEGs) were detected in males (811) than in females (195), and spermatogenesis was the most enriched Gene Ontology (GO) functional classification in males, while circadian regulation of gene expression was the most enriched GO term in females. The expression levels of selected DEGs were routinely verified using qRT-PCR, which showed consistent alterations with the transcriptional changes in RNA-seq data. The causal network analysis by IPA suggested that the adverse outcomes of BPAF in males including liver damage, apoptosis, inflammation of organ, and liver carcinoma, associated with the regulation of several key DEGs detected in RNA-seq, could be linked to the activation of upstream regulatory molecules ifnα, yap1, and ptger2; while, the inhibition of upstream regulators hif1α, ifng, and igf1, leading to the down-regulated expression of several key DEGs, might be involved in BPAF's effects in females. Furthermore, BPAF exposure altered hepatic histological structure and inhibited antioxidant capability in both male and female livers. Overall, this study revealed different regulation networks involved in the sex-dependent effects of BPAF on the fish liver, and these detected DEGs upon BPAF exposure might be used as potential biomarkers for further assessing sex-specific hepatotoxicity following environmental EDC exposure.
双酚AF（BPAF）是一种新兴的内分泌干扰化学物质（EDC），作为双酚A的主要替代品之一在环境中普遍存在。EDC的性别特异性效应已被普遍报道，并且与不同暴露窗口期激素代谢和相关基因表达的性别二态性模式密切相关，但我们对这些机制的理解仍然有限。在此，成年斑马鱼在10μg/L的环境相关浓度BPAF下暴露28天后，分别在雄性和雌性鱼肝中应用RNA测序（RNA-seq）和 Ingenuity通路分析（IPA）研究了全局转录网络，将BPAF长期暴露的性别依赖性毒性与分子反应联系起来。结果，在雄性中检测到的差异表达基因（DEG）（811个）比雌性（195个）更多，精子发生是雄性中最富集的基因本体（GO）功能分类，而基因表达的昼夜节律调节是雌性中最富集的GO术语。使用qRT-PCR常规验证了所选DEG的表达水平，其显示出与RNA-seq数据中的转录变化一致的改变。IPA的因果网络分析表明，BPAF在雄性中的不良后果包括肝损伤、细胞凋亡、器官炎症和肝癌，与RNA-seq中检测到的几个关键DEG的调节相关，可能与上游调节分子ifnα、yap1和ptgerz的激活有关；而上游调节因子hif1α、ifng和igf1的抑制导致几个关键DEG的表达下调，可能参与了BPAF对雌性的影响。此外，BPAF暴露改变了肝组织学结构并抑制了雄性和雌性肝脏中的抗氧化能力。总体而言，本研究揭示了BPAF对鱼肝性别依赖性影响所涉及的不同调节网络，并且这些在BPAF暴露后检测到的DEG可能用作潜在生物标志物，以进一步评估环境EDC暴露后的性别特异性肝毒性。
