Reina-Pérez Iris, Olivas-Martínez Alicia, Mustieles Vicente, Salamanca-Fernández Elena, Molina-Molina José Manuel, Olea Nicolás, Fernández Mariana F
Centre for Biomedical Research & School of Medicine, Radiology and Physical Medicine Department, University of Granada, 18011 Granada, Spain.
Instituto de Investigación Biosanitaria (ibs.GRANADA), 18012 Granada, Spain.
Toxics. 2022 May 27;10(6):287. doi: 10.3390/toxics10060287.
Bisphenol A (BPA) and its substitutes, bisphenol F (BPF) and S (BPS), have previously shown in vitro obesogenic activity. This study was designed to investigate their combined effect on the adipogenic differentiation of human adipose-derived stem cells (hASCs). Cells were exposed for 14 days to an equimolar mixture of bisphenols (MIX) (range 10 nM-10 µM). Oil Red staining was used to measure intracellular lipid accumulation, quantitative real-time polymerase chain reaction (qRT-PCR) to study gene expression of adipogenic markers (PPARγ, C/EBPα, LPL, and FABP4), and Western Blot to determine their corresponding proteins. The MIX promoted intracellular lipid accumulation in a dose-dependent manner with a maximal response at 10 µM. Co-incubation with pure antiestrogen (ICI 182,780) inhibited lipid accumulation, suggesting that the effect was mediated by the estrogen receptor. The MIX also significantly altered the expression of PPARγ, C/EBPα, LPL, and FABP4 markers, observing a non-monotonic (U-shaped) dose-response, with maximal gene expression at 10 nM and 10 µM and lesser expression at 1 µM. This pattern was not observed when bisphenols were tested individually. Exposure to MIX (1-10 µM) also increased all encoded proteins except for FABP4, which showed no changes. Evaluation of the combined effect of relevant chemical mixtures is needed rather than single chemical testing.
双酚A（BPA）及其替代品双酚F（BPF）和双酚S（BPS）先前已显示出体外致肥胖活性。本研究旨在探讨它们对人脂肪来源干细胞（hASCs）成脂分化的联合作用。将细胞暴露于双酚类物质的等摩尔混合物（MIX）（范围为10 nM - 10 µM）中14天。采用油红染色法测量细胞内脂质积累，运用定量实时聚合酶链反应（qRT-PCR）研究成脂标志物（PPARγ、C/EBPα、LPL和FABP4）的基因表达，并通过蛋白质免疫印迹法测定其相应蛋白质。MIX以剂量依赖性方式促进细胞内脂质积累，在10 µM时达到最大反应。与纯抗雌激素药物（ICI 182,780）共同孵育可抑制脂质积累，表明该作用是由雌激素受体介导的。MIX还显著改变了PPARγ、C/EBPα、LPL和FABP4标志物的表达，呈现出非单调（U形）剂量反应，在10 nM和10 µM时基因表达最高，而在1 µM时表达较低。单独测试双酚类物质时未观察到这种模式。暴露于MIX（1 - 10 µM）还增加了除FABP4外的所有编码蛋白，FABP4无变化。需要评估相关化学混合物的联合作用而非单一化学物质测试。
