Cell Biology and Toxicology

, Volume 33, Issue 6, pp 511–526 | Cite as

Comparative microarray analyses of mono(2-ethylhexyl)phthalate impacts on fat cell bioenergetics and adipokine network

  • Huai-Chih Chiang
  • Chih-Hong Wang
  • Szu-Ching Yeh
  • Yi-Hua Lin
  • Ya-Ting Kuo
  • Chih-Wei Liao
  • Feng-Yuan Tsai
  • Wei-Yu Lin
  • Wen-Han Chuang
  • Tsui-Chun Tsou
Original Article


Cellular accumulation of mono(2-ethylhexyl)phthalate (MEHP) has been recently demonstrated to disturb fat cell energy metabolism; however, the underlying mechanism remained unclear. The study aimed to determine how MEHP influenced fat cell transcriptome and how the changes might contribute to bioenergetics. Because of the pivotal role of PPARγ in energy metabolism of fat cells, comparative microarray analysis of gene expression in 3T3-L1 adipocytes treated with both MEHP and rosiglitazone was performed. Pathway enrichment analysis and gene ontology (GO) enrichment analysis revealed that both treatments caused up-regulation of genes involved in PPAR signaling/energy metabolism-related pathways and down-regulation of genes related to adipokine/inflammation signals. MEHP/rosiglitazone-treated adipocytes exhibited increased levels of lipolysis, glucose uptake, and glycolysis; the gene expression profiles provided molecular basis for the functional changes. Moreover, MEHP was shown to induce nuclear translocation and activation of PPARγ. The similarity in gene expression and functional changes in response to MEHP and rosiglitazone suggested that MEHP influenced bioenergetics and adipokine network mainly via PPARγ. Importantly, adipokine levels in C57BL/6J mice with di(2-ethylhexyl)phthalate (DEHP) treatments provided in vivo evidence for microarray results. On the basis of correlation between gene expression and functional assays, possible involvements of genes in bioenergetics of MEHP-treated adipocytes were proposed.


Phthalates Endocrine disruptor Energy metabolism PPARγ Adipocytes 



β-Adrenergic receptor


Database for Annotation, Visualization and Integrated Discovery




Fatty acid-binding proteins


Gene ontology


High-fat diet


Kyoto Encyclopedia of Genes and Genomes




Malonyl-CoA decarboxylase


Normal chow diet


Non-esterified fatty acids


Oxygen consumption rate


Protein Analysis through Evolutionary Relationships


Principal component analysis


Pyruvate dehydrogenase complex


Pyruvate dehydrogenase


PPAR response element


Reduce + Visual Gene Ontology


Type 2 diabetes mellitus


White adipose tissue



This work was supported by grants from the Ministry of Science and Technology (101-2314-B-400-003-MY3, 102-2811-B-400-015, and 103-2811-B-400-022) and the National Health Research Institutes (EO-103-PP-03 and EO-104-PP-03) in Taiwan.

Supplementary material

10565_2016_9380_MOESM1_ESM.docx (84 kb)
ESM 1 (DOCX 84 kb)


  1. Campioli E, Batarseh A, Li J, Papadopoulos V. The endocrine disruptor mono-(2-ethylhexyl) phthalate affects the differentiation of human liposarcoma cells (SW 872). PLoS One. 2011;6:e28750.CrossRefPubMedPubMedCentralGoogle Scholar
  2. Chiang HC, Kuo YT, Shen CC, Lin YH, Wang SL, Tsou TC. Mono(2-ethylhexyl)phthalate accumulation disturbs energy metabolism of fat cells. Arch Toxicol. 2016;90:589–601.CrossRefPubMedGoogle Scholar
  3. Deng T, Sieglaff DH, Zhang A, Lyon CJ, Ayers SD, Cvoro A, et al. A peroxisome proliferator-activated receptor gamma (PPARgamma)/PPARgamma coactivator 1beta autoregulatory loop in adipocyte mitochondrial function. J Biol Chem. 2011;286:30723–31.CrossRefPubMedPubMedCentralGoogle Scholar
  4. Dutchak PA, Katafuchi T, Bookout AL, Choi JH, Yu RT, Mangelsdorf DJ, et al. Fibroblast growth factor-21 regulates PPARgamma activity and the antidiabetic actions of thiazolidinediones. Cell. 2012;148:556–67.CrossRefPubMedPubMedCentralGoogle Scholar
  5. Ellero-Simatos S, Claus SP, Benelli C, Forest C, Letourneur F, Cagnard N, et al. Combined transcriptomic-(1)H NMR metabonomic study reveals that monoethylhexyl phthalate stimulates adipogenesis and glyceroneogenesis in human adipocytes. J Proteome Res. 2011;10:5493–502.CrossRefPubMedPubMedCentralGoogle Scholar
  6. Enguix N, Pardo R, Gonzalez A, Lopez VM, Simo R, Kralli A, et al. Mice lacking PGC-1beta in adipose tissues reveal a dissociation between mitochondrial dysfunction and insulin resistance. Molecular metabolism. 2013;2:215–26.CrossRefPubMedPubMedCentralGoogle Scholar
  7. Feige JN, Gelman L, Rossi D, Zoete V, Metivier R, Tudor C, et al. The endocrine disruptor monoethyl-hexyl-phthalate is a selective peroxisome proliferator-activated receptor gamma modulator that promotes adipogenesis. J Biol Chem. 2007;282:19152–66.CrossRefPubMedGoogle Scholar
  8. Foster DW. Malonyl-CoA: the regulator of fatty acid synthesis and oxidation. J Clin Invest. 2012;122:1958–9.CrossRefPubMedPubMedCentralGoogle Scholar
  9. Furuhashi M, Hotamisligil GS. Fatty acid-binding proteins: role in metabolic diseases and potential as drug targets. Nat Rev Drug Discov. 2008;7:489–503.CrossRefPubMedPubMedCentralGoogle Scholar
  10. Hao C, Cheng X, Xia H, Ma X. The endocrine disruptor mono-(2-ethylhexyl) phthalate promotes adipocyte differentiation and induces obesity in mice. Biosci Rep. 2012;32:619–29.CrossRefPubMedPubMedCentralGoogle Scholar
  11. Hibuse T, Maeda N, Funahashi T, Yamamoto K, Nagasawa A, Mizunoya W, et al. Aquaporin 7 deficiency is associated with development of obesity through activation of adipose glycerol kinase. Proc Natl Acad Sci U S A. 2005;102:10993–8.CrossRefPubMedPubMedCentralGoogle Scholar
  12. Hsu HF, Tsou TC, Chao HR, Kuo YT, Tsai FY, Yeh SC. Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin on adipogenic differentiation and insulin-induced glucose uptake in 3 T3-L1 cells. J Hazard Mater. 2010;182:649–55.CrossRefPubMedGoogle Scholar
  13. James-Todd T, Stahlhut R, Meeker JD, Powell SG, Hauser R, Huang T, et al. Urinary phthalate metabolite concentrations and diabetes among women in the National Health and nutrition examination survey (NHANES) 2001-2008. Environ Health Perspect. 2012;120:1307–13.CrossRefPubMedPubMedCentralGoogle Scholar
  14. Kharitonenkov A, Shiyanova TL, Koester A, Ford AM, Micanovic R, Galbreath EJ, et al. FGF-21 as a novel metabolic regulator. J Clin Invest. 2005;115:1627–35.CrossRefPubMedPubMedCentralGoogle Scholar
  15. Kim JH, Park HY, Bae S, Lim YH, Hong YC. Diethylhexyl phthalates is associated with insulin resistance via oxidative stress in the elderly: a panel study. PLoS One. 2013;8:e71392.CrossRefPubMedPubMedCentralGoogle Scholar
  16. Kleiner S, Mepani RJ, Laznik D, Ye L, Jurczak MJ, Jornayvaz FR, et al. Development of insulin resistance in mice lacking PGC-1alpha in adipose tissues. Proc Natl Acad Sci U S A. 2012;109:9635–40.CrossRefPubMedPubMedCentralGoogle Scholar
  17. Koch HM, Christensen KL, Harth V, Lorber M, Bruning T. Di-n-butyl phthalate (DnBP) and diisobutyl phthalate (DiBP) metabolism in a human volunteer after single oral doses. Arch Toxicol. 2012;86:1829–39.CrossRefPubMedGoogle Scholar
  18. Lee KI, Chiang CW, Lin HC, Zhao JF, Li CT, Shyue SK, et al. Maternal exposure to di-(2-ethylhexyl) phthalate exposure deregulates blood pressure, adiposity, cholesterol metabolism and social interaction in mouse offspring. Archives of toxicology. 2015.Google Scholar
  19. Mehta JL, Li DY. Identification and autoregulation of receptor for OX-LDL in cultured human coronary artery endothelial cells. Biochem Biophys Res Commun. 1998;248:511–4.CrossRefPubMedGoogle Scholar
  20. Mueller E, Drori S, Aiyer A, Yie J, Sarraf P, Chen H, et al. Genetic analysis of adipogenesis through peroxisome proliferator-activated receptor gamma isoforms. J Biol Chem. 2002;277:41925–30.CrossRefPubMedGoogle Scholar
  21. Muzzin P, Eisensmith RC, Copeland KC, Woo SL. Correction of obesity and diabetes in genetically obese mice by leptin gene therapy. Proc Natl Acad Sci U S A. 1996;93:14804–8.CrossRefPubMedPubMedCentralGoogle Scholar
  22. Ouchi N, Parker JL, Lugus JJ, Walsh K. Adipokines in inflammation and metabolic disease. Nat Rev Immunol. 2011;11:85–97.CrossRefPubMedPubMedCentralGoogle Scholar
  23. Palmieri F. The mitochondrial transporter family (SLC25): physiological and pathological implications. Pflugers Archiv: European journal of physiology. 2004;447:689–709.CrossRefPubMedGoogle Scholar
  24. Park BO, Ahrends R, Teruel MN. Consecutive positive feedback loops create a bistable switch that controls preadipocyte-to-adipocyte conversion. Cell Rep. 2012;2:976–90.CrossRefPubMedPubMedCentralGoogle Scholar
  25. Posnack NG, Swift LM, Kay MW, Lee NH, Sarvazyan N. Phthalate exposure changes the metabolic profile of cardiac muscle cells. Environ Health Perspect. 2012;120:1243–51.CrossRefPubMedPubMedCentralGoogle Scholar
  26. Powell E, Kuhn P, Xu W. Nuclear receptor cofactors in PPARgamma-mediated adipogenesis and adipocyte energy metabolism. PPAR Res. 2007;2007:53843.CrossRefPubMedGoogle Scholar
  27. Samuel P, Khan MA, Nag S, Inagami T, Hussain T. Angiotensin AT(2) receptor contributes towards gender bias in weight gain. PLoS One. 2013;8:e48425.CrossRefPubMedPubMedCentralGoogle Scholar
  28. Savage DB. PPAR gamma as a metabolic regulator: insights from genomics and pharmacology. Expert reviews in molecular medicine. 2005;7:1–16.CrossRefPubMedGoogle Scholar
  29. Sharma AM, Staels B. Review: peroxisome proliferator-activated receptor gamma and adipose tissue--understanding obesity-related changes in regulation of lipid and glucose metabolism. J Clin Endocrinol Metab. 2007;92:386–95.CrossRefPubMedGoogle Scholar
  30. Soni KG, Lehner R, Metalnikov P, O'Donnell P, Semache M, Gao W, et al. Carboxylesterase 3 (EC is a major adipocyte lipase. J Biol Chem. 2004;279:40683–9.CrossRefPubMedGoogle Scholar
  31. Spalding KL, Arner E, Westermark PO, Bernard S, Buchholz BA, Bergmann O, et al. Dynamics of fat cell turnover in humans. Nature. 2008;453:783–7.CrossRefPubMedGoogle Scholar
  32. Tsai FY, Cheng YT, Tsou TC. A recombinant PPRE-driven luciferase bioassay for identification of potential PPAR agonists. Vitam Horm. 2014;94:427–35.CrossRefPubMedGoogle Scholar
  33. Vega RB, Huss JM, Kelly DP. The coactivator PGC-1 cooperates with peroxisome proliferator-activated receptor alpha in transcriptional control of nuclear genes encoding mitochondrial fatty acid oxidation enzymes. Mol Cell Biol. 2000;20:1868–76.CrossRefPubMedPubMedCentralGoogle Scholar
  34. Wang H, Zhou Y, Tang C, He Y, Wu J, Chen Y, et al. Urinary phthalate metabolites are associated with body mass index and waist circumference in Chinese school children. PLoS One. 2013;8:e56800.CrossRefPubMedPubMedCentralGoogle Scholar
  35. Wang P, Renes J, Bouwman F, Bunschoten A, Mariman E, Keijer J. Absence of an adipogenic effect of rosiglitazone on mature 3 T3-L1 adipocytes: increase of lipid catabolism and reduction of adipokine expression. Diabetologia. 2007;50:654–65.CrossRefPubMedPubMedCentralGoogle Scholar
  36. Xu A, Lam MC, Chan KW, Wang Y, Zhang J, Hoo RL, et al. Angiopoietin-like protein 4 decreases blood glucose and improves glucose tolerance but induces hyperlipidemia and hepatic steatosis in mice. Proc Natl Acad Sci U S A. 2005;102:6086–91.CrossRefPubMedPubMedCentralGoogle Scholar
  37. Zeng Q, Wei C, Wu Y, Li K, Ding S, Yuan J, et al. Approach to distribution and accumulation of dibutyl phthalate in rats by immunoassay. Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association. 2013;56:18–27.CrossRefGoogle Scholar
  38. Zhang S, Hulver MW, McMillan RP, Cline MA, Gilbert ER. The pivotal role of pyruvate dehydrogenase kinases in metabolic flexibility. Nutrition & metabolism. 2014;11:10.CrossRefGoogle Scholar
  39. Zhang YH, Chen BH, Zheng LX, Wu XY. Study on the level of phthalates in human biological samples. Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine]. 2003;37:429–34.Google Scholar
  40. Zhao JF, Hsiao SH, Hsu MH, Pao KC, Kou YR, Shyue SK, et al. Di-(2-ethylhexyl) phthalate accelerates atherosclerosis in apolipoprotein E-deficient mice. Archives of toxicology. 2014.Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  1. 1.National Institute of Environmental Health SciencesNational Health Research InstitutesZhunanTaiwan
  2. 2.Department of Biological Science and TechnologyNational Chiao Tung UniversityHsinchuTaiwan

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