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Differential effects on adiposity and serum marker of bone formation by post-weaning exposure to methylparaben and butylparaben


Paraben esters and their salts are widely used as preservatives in cosmetics, personal care products, pharmaceuticals, and foods. We and others have reported that parabens promote adipogenesis in vitro. Here, we investigated the effects of post-weaning exposure to parabens (methylparaben and butylparaben) on body weight, white adipose tissue mass, and obesity associated metabolic biomarkers in female obesity-prone C57BL/6J mice fed with a chow diet or a high fat diet. Methylparaben exposure by daily oral gavage (100 mg/kg/day) increased adiposity and serum leptin levels compared to the controls when fed the chow diet, but not the high fat diet. In contrast, butylparaben exposure did not induce such effects. Exposure to either paraben induced changes in gene expression related to adipocyte differentiation and lipogenesis in the white adipose tissue (WAT) and the liver, regardless of diet. Moreover, exposure to both parabens under the chow diet significantly decreased serum procollagen type 1 N-terminal propeptide (P1NP) but had no effects on C-terminal telopeptide of type I collagen (CTX-I) levels, suggesting that post-weaning exposure to paraben may negatively affect bone formation, but not bone resorption. Taken together, our results demonstrate that post-weaning exposure to paraben, methylparaben in particular, promotes adipogenesis but suppresses serum marker of bone formation in vivo. Our results add to the growing body of literature indicating potential negative health outcomes associated with paraben exposure. Further study of early life exposure to paraben on the development of fat and bone is warranted.

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Adipose differentiation-related protein


Area under curve


Brown adipose tissue


CCAAT/enhancer binding protein alpha


Endocrine disrupting chemical


Collagen type I C-terminal telopeptide


Fatty acid binding protein 4


Fatty acid synthase


Glucose tolerance test


Insulin tolerance test


Procollagen type 1 N-terminal propeptide




Peroxisome proliferator-activated receptor gamma






Stearoyl CoA desaturase




White adipose tissue


  1. Abdallah BM, Kassem M (2012) New factors controlling the balance between osteoblastogenesis and adipogenesis. Bone 50:540–545

  2. Ayala JE et al. (2010) Standard operating procedures for describing and performing metabolic tests of glucose homeostasis in mice. Dis Model Mech 3:525–534

  3. Barouki R, Gluckman PD, Grandjean P, Hanson M, Heindel JJ (2012) Developmental origins of non-communicable disease: implications for research and public health Environ Health 11:42

  4. Beausoleil C et al. (2013) Low dose effects and non-monotonic dose responses for endocrine active chemicals: science to practice workshop: workshop summary. Chemosphere 93:847–856

  5. Bledzka D, Gromadzinska J, Wasowicz W (2014) Parabens. From environmental studies to human health. Environ Int 67:27–42

  6. Calafat AM et al. (2009) Exposure to bisphenol A and other phenols in neonatal intensive care unit premature infants. Environ Health Perspect 117:639–644

  7. Calafat AM, Ye X, Wong LY, Bishop AM, Needham LL (2010) Urinary concentrations of four parabens in the U.S. population: NHANES 2005–2006. Environ Health Perspect 118:679–685

  8. Chen Q et al. (2016) Fate decision of mesenchymal stem cells: adipocytes or osteoblasts? Cell Death Differ. doi:10.1038/cdd.2015.168

  9. Cosmetic Ingredient Review (CIR) (2008) Final amended report on the safety assessment of methylparaben, ethylparaben, propylparaben, isopropylparaben, butylparaben, isobutylparaben, and benzylparaben as used in cosmetic products. Int J Toxicol 27(Suppl 4):1–82

  10. Frederiksen H, Taxvig C, Hass U, Vinggaard AM, Nellemann C (2008) Higher levels of ethylparaben and butylparaben in rat amniotic fluid than in maternal plasma after subcutaneous administration. Toxicol Sci : Off J Soc Toxicol 106:376–383

  11. Frederiksen H, Jorgensen N, Andersson AM (2011) Parabens in urine, serum and seminal plasma from healthy Danish men determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). J Expo Sci Environ Epidemiol 21:262–271

  12. Garcia-Arevalo M, Alonso-Magdalena P, Rebelo Dos Santos J, Quesada I, Carneiro EM, Nadal A (2014) Exposure to bisphenol-A during pregnancy partially mimics the effects of a high-fat diet altering glucose homeostasis and gene expression in adult male mice. PLoS One 9:e100214

  13. Gesta S, Tseng YH, Kahn CR (2007) Developmental origin of fat: tracking obesity to its source. Cell 131:242–256

  14. Glatt V, Canalis E, Stadmeyer L, Bouxsein ML (2007) Age-related changes in trabecular architecture differ in female and male C57BL/6J mice. J Bone Miner Res Off J Am Soc Bone Miner Res 22:1197–1207

  15. Gomez E et al. (2005) Estrogenic activity of cosmetic components in reporter cell lines: parabens, UV screens, and musks. J Toxicol Environ Health Part A 68:239–251

  16. Gosman JH, Stout SD, Larsen CS (2011) Skeletal biology over the life span: a view from the surfaces. Am J Phys Anthropol 146(Suppl 53):86–98

  17. Guo Y, Kannan K (2013) A survey of phthalates and parabens in personal care products from the United States and its implications for human exposure. Environ Sci Technol 47:14442–14449

  18. Halloran BP, Ferguson VL, Simske SJ, Burghardt A, Venton LL, Majumdar S (2002) Changes in bone structure and mass with advancing age in the male C57BL/6J mouse. J Bone Miner Res Off J Am Soc Bone Miner Res 17:1044–1050

  19. Harville HM, Voorman R, Prusakiewicz JJ (2007) Comparison of paraben stability in human and rat skin. Drug Metab Lett 1:17–21

  20. Heindel JJ et al. (2015) Developmental origins of health and disease: integrating environmental influences. Endocrinology 156:3416–3421

  21. Hu P et al. (2013) Effects of parabens on adipocyte differentiation. Toxicol Sci Off J Soc Toxicol 131:56–70

  22. Imai T, Taketani M, Shii M, Hosokawa M, Chiba K (2006) Substrate specificity of carboxylesterase isozymes and their contribution to hydrolase activity in human liver and small intestine. Drug Metab Dispos 34:1734–1741

  23. Ishiwatari S, Suzuki T, Hitomi T, Yoshino T, Matsukuma S, Tsuji T (2007) Effects of methyl paraben on skin keratinocytes. J Appl Toxicol JAT 27:1–9

  24. James AW (2013) Review of signaling pathways governing MSC osteogenic and adipogenic differentiation. Scientifica (Cairo) 2013:684736

  25. Janjua NR, Frederiksen H, Skakkebaek NE, Wulf HC, Andersson AM (2008) Urinary excretion of phthalates and paraben after repeated whole-body topical application in humans. Int J Androl 31:118–130

  26. Jimenez-Diaz I et al. (2011) A new liquid chromatography-tandem mass spectrometry method for determination of parabens in human placental tissue samples. Talanta 84:702–709

  27. Karpuzoglu E, Holladay SD, Gogal RM Jr (2013) Parabens: potential impact of low-affinity estrogen receptor binding chemicals on human health. J Toxicol Environ Health B Crit Rev 16:321–335

  28. Khanna S, Darbre PD (2013) Parabens enable suspension growth of MCF-10A immortalized, non-transformed human breast epithelial cells. J Appl Toxicol JAT 33:378–382

  29. Kirchner S, Kieu T, Chow C, Casey S, Blumberg B (2010) Prenatal exposure to the environmental obesogen tributyltin predisposes multipotent stem cells to become adipocytes. Mol Endocrinol 24:526–539

  30. Kovacs CS (2011) Bone development in the fetus and neonate: role of the calciotropic hormones. Curr Osteoporos Rep 9:274–283

  31. Lagarde F, Beausoleil C, Belcher SM, Belzunces LP, Emond C, Guerbet M, Rousselle C (2015) Non-monotonic dose-response relationships and endocrine disruptors: a qualitative method of assessment Environ Health 14:13 doi:10.1186/1476-069X-14-13

  32. Lemini C, Jaimez R, Avila ME, Franco Y, Larrea F, Lemus AE (2003) In vivo and in vitro estrogen bioactivities of alkyl parabens. Toxicol Ind Health 19:69–79

  33. Liao C, Liu F, Kannan K (2013) Occurrence of and dietary exposure to parabens in foodstuffs from the United States. Environ Sci Technol 47:3918–3925

  34. Moon MK et al. (2015) Long-term oral exposure to bisphenol A induces glucose intolerance and insulin resistance. J Endocrinol 226:35–42

  35. Naville D, Labaronne E, Vega N, Pinteur C, Canet-Soulas E, Vidal H, Le Magueresse-Battistoni B (2015) Metabolic outcome of female mice exposed to a mixture of low-dose pollutants in a diet-induced obesity model. PLoS One 10:e0124015

  36. Nishikawa S, Yasoshima A, Doi K, Nakayama H, Uetsuka K (2007) Involvement of sex, strain and age factors in high fat diet-induced obesity in C57BL/6J and BALB/cA mice. Exp Anim 56:263–272

  37. Oishi S (2001) Effects of butylparaben on the male reproductive system in rats. Toxicol Ind Health 17:31–39

  38. Oishi S (2002a) Effects of butyl paraben on the male reproductive system in mice. Arch Toxicol 76:423–429

  39. Oishi S (2002b) Effects of propyl paraben on the male reproductive system. Food Chem Toxicol Int J Publ Br Ind Biol Res Assoc 40:1807–1813

  40. Oishi S (2004) Lack of spermatotoxic effects of methyl and ethyl esters of p-hydroxybenzoic acid in rats. Food Chem Toxicol Int J Publ Br Ind Biol Res Assoc 42:1845–1849

  41. Ozaki H et al. (2013) Comparative study of the hydrolytic metabolism of methyl-, ethyl-, propyl-, butyl-, heptyl- and dodecylparaben by microsomes of various rat and human tissues. Xenobiotica 43:1064–1072

  42. Pereira-Fernandes A, Demaegdt H, Vandermeiren K, Hectors TL, Jorens PG, Blust R, Vanparys C (2013) Evaluation of a screening system for obesogenic compounds: screening of endocrine disrupting compounds and evaluation of the PPAR dependency of the effect PLoS One 8:e77481

  43. Philippat C, Bennett D, Calafat AM, Picciotto IH (2015) Exposure to select phthalates and phenols through use of personal care products among Californian adults and their children. Environ Res 140:369–376

  44. Poissonnet CM, Burdi AR, Garn SM (1984) The chronology of adipose tissue appearance and distribution in the human fetus. Early Hum Dev 10:1–11

  45. Prusakiewicz JJ, Ackermann C, Voorman R (2006) Comparison of skin esterase activities from different species. Pharm Res 23:1517–1524

  46. Pycke BF, Geer LA, Dalloul M, Abulafia O, Halden RU (2015) Maternal and fetal exposure to parabens in a multiethnic urban U.S. population. Environ Int 84:193–200

  47. SCCP/1017/06 (2006) The Scientific Committee on Consumer Products (SCCP) opinion on parabens (Colipa no. P82), adopted during the 9th plenary meeting of 10 October 2006

  48. Schlumpf M et al. (2010) Exposure patterns of UV filters, fragrances, parabens, phthalates, organochlor pesticides, PBDEs, and PCBs in human milk: correlation of UV filters with use of cosmetics. Chemosphere 81:1171–1183

  49. Taher L, Collette NM, Murugesh D, Maxwell E, Ovcharenko I, Loots GG (2011) Global gene expression analysis of murine limb development PLoS One 6:e28358

  50. Teitelbaum SL et al. (2016) Paired serum and urine concentrations of biomarkers of diethyl phthalate, methylparaben, and triclosan in rats. Environ Health Perspect 124:39–45

  51. van Esterik JC, Dolle ME, Lamoree MH, van Leeuwen SP, Hamers T, Legler J, van der Ven LT (2014) Programming of metabolic effects in C57BL/6JxFVB mice by exposure to bisphenol A during gestation and lactation. Toxicology 321:40–52

  52. Vandenberg LN (2014) Non-monotonic dose responses in studies of endocrine disrupting chemicals: bisphenol a as a case study. Dose-Response 12:259–276

  53. Watt J, Schlezinger JJ (2015) Structurally-diverse, PPARgamma-activating environmental toxicants induce adipogenesis and suppress osteogenesis in bone marrow mesenchymal stromal cells. Toxicology 331:66–77

  54. Yang M et al. (2016) Bisphenol A promotes adiposity and inflammation in a nonmonotonic dose-response way in 5-week-old male and female C57BL/6J mice fed a low-calorie diet. Endocrinology 157:2333–2345

  55. Ye X, Bishop AM, Reidy JA, Needham LL, Calafat AM (2006) Parabens as urinary biomarkers of exposure in humans. Environ Health Perspect 114:1843–1846

  56. Ye X, Wong LY, Jia LT, Needham LL, Calafat AM (2009) Stability of the conjugated species of environmental phenols and parabens in human serum. Environ Int 35:1160–1163

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We thank Dr. Huanbiao Mo for critically reading the manuscript. The studies were supported by the faculty start-up fund (L.Z.) and in part by NIEHS 1R21ES017475-01A1 (J.C.). The funding sources had no roles in the study design; the collection, analysis, and interpretation of data; the writing of the report; and the decision to submit the article for publication.

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Correspondence to Ling Zhao.

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All animal procedures and protocols were approved by the Institutional Animal Care and Use Committee at the University of Tennessee, Knoxville. This investigation was conducted in an animal facility fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care.

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The authors declare that they have no conflict of interest.

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Responsible editor: Philippe Garrigues

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Hu, P., Kennedy, R.C., Chen, X. et al. Differential effects on adiposity and serum marker of bone formation by post-weaning exposure to methylparaben and butylparaben. Environ Sci Pollut Res 23, 21957–21968 (2016).

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  • Paraben
  • Endocrine disruptor
  • Adipogenesis
  • Bone formation
  • P1NP
  • CTX-I