Watkins Andrew M, Wood Carmen R, Lin Mimi T, Abbott Barbara D
Developmental Toxicology Branch, Toxicity Assessment Division, National Health and Environmental Effects Research Lab, Office of Research and Development, US Environmental Protection Agency, Research Triangle Park, NC 27709, USA.
Developmental Toxicology Branch, Toxicity Assessment Division, National Health and Environmental Effects Research Lab, Office of Research and Development, US Environmental Protection Agency, Research Triangle Park, NC 27709, USA.
Mol Cell Endocrinol. 2015 Jan 15;400:90-101. doi: 10.1016/j.mce.2014.10.020. Epub 2014 Nov 5.
The 3T3-L1 preadipocyte culture system has been used to examine numerous compounds that influence adipocyte differentiation or function. The perfluoroalkyl acids (PFAAs), used as surfactants in a variety of industrial applications, are of concern as environmental contaminants that are detected worldwide in human serum and animal tissues. This study was designed to evaluate the potential for PFAAs to affect adipocyte differentiation and lipid accumulation using mouse 3T3-L1 cells. Cells were treated with perfluorooctanoic acid (PFOA) (5-100 µM), perfluorononanoic acid (PFNA) (5-100 µM), perfluorooctane sulfonate (PFOS) (50-300 µM), perfluorohexane sulfonate (PFHxS) (40-250 µM), the peroxisome proliferator activated receptor (PPAR) PPARα agonist Wyeth-14,643 (WY-14,643), and the PPARγ agonist rosiglitazone. The PPARγ agonist was included as a positive control as this pathway is critical to adipocyte differentiation. The PPARα agonist was included as the PFAA compounds are known activators of this pathway. Cells were assessed morphometrically and biochemically for number, size, and lipid content. RNA was extracted for qPCR analysis of 13 genes selected for their importance in adipocyte differentiation and lipid metabolism. There was a significant concentration-related increase in cell number and decreased cell size after exposure to PFOA, PFHxS, PFOS, and PFNA. All four PFAA treatments produced a concentration-related decrease in the calculated average area occupied by lipid per cell. However, total triglyceride levels per well increased with a concentration-related trend for all compounds, likely due to the increased cell number. Expression of mRNA for the selected genes was affected by all exposures and the specific impacts depended on the particular compound and concentration. Acox1 and Gapdh were upregulated by all six compounds. The strongest overall effect was a nearly 10-fold induction of Scd1 by PFHxS. The sulfonated PFAAs produced numerous, strong changes in gene expression similar to the effects after treatment with the PPARγ agonist rosiglitazone. By comparison, the effects on gene expression were muted for the carboxylated PFAAs and for the PPARα agonist WY-14,643. In summary, all perfluorinated compounds increased cell number, decreased cell size, increased total triglyceride, and altered expression of genes associated with adipocyte differentiation and lipid metabolism.
3T3-L1前脂肪细胞培养系统已被用于检测众多影响脂肪细胞分化或功能的化合物。全氟烷基酸（PFAAs）在各种工业应用中用作表面活性剂，作为环境污染物受到关注，在全球范围内的人体血清和动物组织中均可检测到。本研究旨在使用小鼠3T3-L1细胞评估PFAAs影响脂肪细胞分化和脂质积累的可能性。细胞分别用全氟辛酸（PFOA）（5-100µM）、全氟壬酸（PFNA）（5-100µM）、全氟辛烷磺酸（PFOS）（50-300µM）、全氟己烷磺酸（PFHxS）（40-250µM）、过氧化物酶体增殖物激活受体（PPAR）PPARα激动剂Wyeth-14,643（WY-14,643）以及PPARγ激动剂罗格列酮进行处理。将PPARγ激动剂作为阳性对照，因为该途径对脂肪细胞分化至关重要。将PPARα激动剂纳入是因为PFAA化合物是该途径已知的激活剂。对细胞进行形态学和生化评估，以确定细胞数量、大小和脂质含量。提取RNA用于对13个因其在脂肪细胞分化和脂质代谢中的重要性而选择的基因进行qPCR分析。暴露于PFOA、PFHxS、PFOS和PFNA后，细胞数量显著呈浓度相关增加，细胞大小减小。所有四种PFAA处理均使计算得出的每个细胞脂质占据的平均面积呈浓度相关减少。然而，所有化合物每孔总甘油三酯水平均呈浓度相关趋势增加，这可能是由于细胞数量增加所致。所选基因的mRNA表达受所有暴露影响，具体影响取决于特定化合物和浓度。六种化合物均使Acox1和Gapdh上调。总体影响最强的是PFHxS使Scd1诱导近10倍。磺化PFAAs在基因表达上产生了许多强烈变化，类似于用PPARγ激动剂罗格列酮处理后的效果。相比之下，羧化PFAAs和PPARα激动剂WY-14,643对基因表达的影响较小。总之，所有全氟化合物均增加细胞数量、减小细胞大小、增加总甘油三酯，并改变与脂肪细胞分化和脂质代谢相关的基因表达。
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