RECETOX, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic.
Environ Sci Technol. 2023 Feb 7;57(5):2006-2018. doi: 10.1021/acs.est.2c03998. Epub 2023 Jan 24.
The present study aims to evaluate the effects of repeated exposure to 2-ethylhexyldiphenyl phosphate (EHDPP) on human liver cells. three-dimensional (3D) hepatospheroid cell culture was utilized to explore the potential mechanisms of EHDPP-mediated metabolic disruption through morphological, transcriptional, and biochemical assays. Lipidomics analysis was performed on the individual hepatospheroids to investigate the effects on intracellular lipid profiles, followed by hepatospheroid morphology, growth, functional parameters, and cytotoxicity evaluation. The possible mechanisms were delineated using the gene-level analysis by assessing the expression of key genes encoding for hepatic lipid metabolism. We revealed that exposure to EHDPP at 1 and 10 μM for 7 days alters the lipid profile of human 3D hepatospheroids. Dysregulation in several lipid classes, including sterol lipids (cholesterol esters), sphingolipids (dihydroceramide, hexosylceramide, ceramide, sphingomyelin), glycerolipids (triglycerides), glycerophospholipids, and fatty acyls, was noted along with alteration in genes including ACAT1, ACAT2, CYP27A1, ABCA1, GPAT2, PNPLA2, PGC1α, and Nrf2. Our study brings a novel insight into the metabolic disrupting effects of EHDPP and demonstrates the utility of hepatospheroids as an cell culture model complemented with omics technology (, lipidomics) for mechanistic toxicity studies.
本研究旨在评估重复接触 2-乙基己基二苯基磷酸酯 (EHDPP) 对人肝细胞的影响。采用三维（3D）肝球体细胞培养技术，通过形态学、转录组和生化分析探讨 EHDPP 介导的代谢紊乱的潜在机制。对单个肝球体进行脂质组学分析，以研究其对细胞内脂质谱的影响，随后评估肝球体的形态、生长、功能参数和细胞毒性。通过评估编码肝脂代谢的关键基因的表达，利用基因水平分析来描述可能的机制。结果表明，在 7 天内以 1 和 10 μM 的浓度暴露于 EHDPP 会改变人 3D 肝球体的脂质谱。几种脂质类别的失调，包括固醇脂质（胆固醇酯）、鞘脂（二氢神经酰胺、己糖神经酰胺、神经酰胺、神经鞘磷脂）、甘油脂质（甘油三酯）、甘油磷酸脂质和脂肪酸酰基，以及包括 ACAT1、ACAT2、CYP27A1、ABCA1、GPAT2、PNPLA2、PGC1α 和 Nrf2 在内的基因的改变都有被注意到。本研究为 EHDPP 的代谢干扰效应提供了新的见解，并展示了肝球体作为细胞培养模型与组学技术（如脂质组学）相结合用于机制毒性研究的应用。
