Di-(2-ethylhexyl) phthalate induces precocious puberty in adolescent female rats

Document Type: Original Article


1 Scientific Research Center, China-Japan Union Hospital of Jilin University, Changchun, 130033, China

2 Department of Orthodontics, School and Hospital of Stomatology, Jilin University, Changchun, 130021, China

3 Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun 130033, China

4 Scientific Research Center, China-Japan Union Hospital of Jilin University, Changchun, China.

5 Department of Dermatological, China-Japan Union Hospital of Jilin University, Changchun, 130033, China

6 Department of Endocrinology, China-Japan Union Hospital of Jilin University, Changchun, 130033, China


Objective(s): Nowadays, Di-(2-ethylhexyl) phthalate (DEHP) is widely used in different kinds of commercial products as a plasticizer. Previous studies have revealed that exposures to DEHP could be associated with precocious puberty in teenagers, but the exact mechanism is yet to be known.
Materials and Methods: In this study, 48 prepubertal Wistar female rats were randomly apportioned into 4 groups and orally treated with 0, 250, 500, and 1000 mg/kg/d DEHP from postnatal day 21 up to 4 weeks. Subsequently, we examined the indicators related to the initiation of sexual development.
Results: DEHP was able to shorten the vaginal opening time and prolong the estrous cycles of female rats. IGF-1 expression was significantly upregulated by 1000 mg/kg/d DEHP in the hypothalamus, and the hypothalamic, as well as serum levels of GH, were also upregulated by DEHP. It also caused decrements in serum levels of FSH, LH, and T and the increment in level of progesterone. Meanwhile, DEHP was able to exert its effect on the mRNA and protein expression levels of Kiss-1, GPR54, and GnRH in the hypothalamus in pubertal female rats.
Conclusion: These findings are revealing that DEHP exposure more likely causes imbalances of hypothalamus functioning in pubertal female rats and thus induces precautious puberty in these animals.


Main Subjects

1. Macedo DB, Silveira LF, Bessa DS, Brito VN, Latronico AC. Sexual precocity--genetic bases of central precocious puberty and autonomous gonadal activation. Endocr Dev 2016; 29:50-71.
2. Ozen S, Darcan S. Effects of environmental endocrine disruptors on pubertal development. J Clin Res Pediatr Endocrinol 2011; 3:1-6.
3. Gore AC, Chappell VA, Fenton SE, Flaws JA, Nadal A, Prins GS, et al. EDC-2: the endocrine society's second scientific statement on endocrine-disrupting chemicals. Endocr Rev 2015; 36:E1-E150.
4. Meeker JD. Exposure to environmental endocrine disruptors and child development. Arch Pediatr Adolesc Med 2012; 166:E1-7.
5. Behie AM, O'Donnell MH. Prenatal smoking and age at menarche: influence of the prenatal environment on the timing of puberty. Hum Reprod 2015; 30:957-962.
6. Maqbool F, Mostafalou S, Bahadar H, Abdollahi M. Review of endocrine disorders associated with environmental toxicants and possible involved mechanisms. Life Sci 2016; 145:265-273.
7. Wolff MS, Teitelbaum SL, Pinney SM, Windham G, Liao L, Biro F, et al. Investigation of relationships between urinary biomarkers of phytoestrogens, phthalates, and phenols and pubertal stages in girls. Environ Health Perspect 2010; 118:1039-1046.
8. Gee RH, Charles A, Taylor N, Darbre PD. Oestrogenic and androgenic activity of triclosan in breast cancer cells. J Appl Toxicol 2008; 28:78-91.
9. Magdouli S, Daghrir R, Brar SK, Drogui P, Tyagi RD. Di 2-ethylhexylphtalate in the aquatic and terrestrial environment: a critical review. J Environ Manage 2013; 127:36-49.
10. Zolfaghari M, Drogui P, Seyhi B, Brar SK, Buelna G, Dube R. Occurrence, fate and effects of Di (2-ethylhexyl) Phthalate in wastewater treatment plants: a review. Environ Pollut 2014; 194:281-293.
11. Song M, Chi C, Guo M, Wang X, Cheng L, Shen X. Pollution levels and characteristics of phthalate esters in indoor air of offices. J Environ Sci 2015; 28:157-162.
12. Lovekamp-Swan T, Davis BJ. Mechanisms of phthalate ester toxicity in the female reproductive system. Environ Health Perspect 2003; 111:139-145.
13. Xia X, Yang L, Bu Q, Liu R. Levels, distribution, and health risk of phthalate esters in urban soils of Beijing, China. J Environ Qual 2011; 40:1643-1651.
14. Liu N, Wang Y, Yang Q, Lv Y, Jin X, Giesy JP, et al. Probabilistic assessment of risks of diethylhexyl phthalate (DEHP) in surface waters of China on reproduction of fish. Environ Pollut 2016; 213:482-488.
15. Somasundaram DB, Selvanesan BC, Ramachandran I, Bhaskaran RS. Lactational exposure to di(2-ethylhexyl) phthalate impairs the ovarian and uterine function of adult offspring rat. Reprod Sci 2016; 23:549-559.
16. Pocar P, Fiandanese N, Secchi C, Berrini A, Fischer B, Schmidt JS, et al. Exposure to di(2-ethyl-hexyl) phthalate (DEHP) in utero and during lactation causes long-term pituitary-gonadal axis disruption in male and female mouse offspring. Endocrinology 2012; 153:937-948.
17. Vickers MH. Developmental programming and adult obesity: the role of leptin. Curr Opin Endocrinol Diabetes Obes 2007; 14:17-22.
18. Xi W, Wan HT, Zhao YG, Wong MH, Giesy JP, Wong CK. Effects of perinatal exposure to bisphenol A and di(2-ethylhexyl) phthalate on gonadal development of male mice. Environ Sci Pollut Res Int 2011; 19:2515-2527.
19. Noroozi A, Shirazi MR, Zamiri MJ, Tamadon A, Akhlaghi A, Tanideh N, et al. Increased litter size and suckling intensity inhibit KiSS-1 mRNA expression in rat arcuate nucleus. Iran J Basic Med Sci 2014; 17:600-605.
20. Li SN, Xue HL, Zhang Q, Xu JH, Wang S, Chen L, et al. Photoperiod regulates the differential expression of KiSS-1 and GPR54 in various tissues and sexes of striped hamster. Genet Mol Res 2015; 14:13894-13905.
21. Rousseau-Nepton I, Kaduri S, Garfield N, Krishnamoorthy P. Hypothalamic hamartoma associated with central precocious puberty and growth hormone deficiency. J Pediatr Endocrinol Metab 2014; 27:117-121.
22. He Y, Sun W, Yu J. Is precocious puberty linked to hypothalamic expression of arginine-phenylalanine-amide-related peptide?. Iran J Basic Med Sci 2017; 20:1074-1078.
23. Bulus AD, Asci A, Erkekoglu P, Balci A, Andiran N, Kocer-Gumusel B. The evaluation of possible role of endocrine disruptors in central and peripheral precocious puberty. Toxicol Mech Methods 2016; 26:493-500.
24. Naseri M, Parham A, Moghimi A. The effect of sodium thiopental as a GABA mimetic drug in neonatal period on expression of GAD65 and GAD67 genes in hippocampus of newborn and adult male rats. Iran J Basic Med Sci 2017; 20:996-1001.
25. Wen Y, Liu SD, Lei X, Ling YS, Luo Y, Liu Q. Association of PAEs with precocious puberty in children: a systematic review and meta-analysis. Int J Environ Res Public Health 2015; 12:15254-15268.
26. Li X, Cai D. Single and combined toxic effects of di-2-ethylhexyl phthalate and cypermethrin on fertility and development in the the prepubertal male rats. Wei Sheng Yan Jiu 2012; 41:710-716.
27. Kavlock R, Boekelheide K, Chapin R, Cunningham M, Faustman E, Foster P, et al. NTP center for the evaluation of risks to human reproduction: phthalates expert panel report on the reproductive and developmental toxicity of di-n-butyl phthalate. Reprod Toxicol 2002; 16:489-527.
28. Heudorf U, Mersch-Sundermann V, Angerer J. Phthalates: toxicology and exposure. Int J Hyg Environ Health 2007; 210:623-634.
29. Deutschle T, Reiter R, Butte W, Heinzow B, Keck T, Riechelmann H. A controlled challenge study on di(2-ethylhexyl) phthalate (DEHP) in house dust and the immune response in human nasal mucosa of allergic subjects. Environ Health Perspect 2008; 116:1487-1493.
30. Carbone S, Samaniego YA, Cutrera R, Reynoso R, Cardoso N, Scacchi P, et al. Different effects by sex on hypothalamic-pituitary axis of prepubertal offspring rats produced by in utero and lactational exposure to di-(2-ethylhexyl) phthalate (DEHP). Neurotoxicology 2012; 33:78-84.
31. Xu Y, Agrawal S, Cook TJ, Knipp GT. Di-(2-ethylhexyl)-phthalate affects lipid profiling in fetal rat brain upon maternal exposure. Arch Toxicol 2007; 81:57-62.
32. Liu T, Li N, Zhu J, Yu G, Guo K, Zhou L, et al. Effects of di-(2-ethylhexyl) phthalate on the hypothalamus-pituitary-ovarian axis in adult female rats. Reprod Toxicol 2014; 46:141-147.
33. Liu T, Jia Y, Zhou L, Wang Q, Sun D, Xu J, et al. Effects of di-(2-ethylhexyl) phthalate on the hypothalamus-uterus in pubertal female rats. Int J Environ Res Public Health 2016; 13:E1130.
34. Costa EM, Spritzer PM, Hohl A, Bachega TA. Effects of endocrine disruptors in the development of the female reproductive tract. Arq Bras Endocrinol Metabol 2014; 58:153-161.
35. Ashpole NM, Sanders JE, Hodges EL, Yan H, Sonntag WE. Growth hormone, insulin-like growth factor-1 and the aging brain. Exp Gerontol 2015; 68:76-81.
36. Habibi P, Babri S, Ahmadiasl N, Yousefi H. Effects of genistein and swimming exercise on spatial memory and expression of microRNA 132, BDNF, and IGF-1 genes in the hippocampus of ovariectomized rats. Iran J Basic Med Sci 2017; 20:856-862.
37. Huang HB, Pan WH, Chang JW, Chiang HC, Guo YL, Jaakkola JJ, et al. Does exposure to phthalates influence thyroid function and growth hormone homeostasis? The taiwan environmental survey for toxicants (TEST) 2013. Environ Res 2017; 153:63-72.
38. Silva MJ, Barr DB, Reidy JA, Malek NA, Hodge CC, Caudill SP, et al. Urinary levels of seven phthalate metabolites in the U.S. population from the national health and nutrition examination survey (NHANES) 1999-2000. Environ Health Perspect 2004; 112:331-338.
39. Mayer C, Boehm U. Female reproductive maturation in the absence of kisspeptin/GPR54 signaling. Nat Neurosci 2011; 14:704-710.
40. Navarro VM. New insights into the control of pulsatile GnRH release: the role of Kiss1/neurokinin B neurons. Front Endocrinol(Lausanne) 2012; 3:48.
41. Israel DD, Sheffer-Babila S, de Luca C, Jo YH, Liu SM, Xia Q, et al. Effects of leptin and melanocortin signaling interactions on pubertal development and reproduction. Endocrinology 2012; 153:2408-2419.
42. Nakane R, Oka Y. Excitatory action of GABA in the terminal nerve gonadotropin-releasing hormone neurons. J Neurophysiol 2010; 103:1375-1384.
43. Bellefontaine N, Hanchate NK, Parkash J, Campagne C, de Seranno S, Clasadonte J, et al. Nitric oxide as key mediator of neuron-to-neuron and endothelia-to-glia communication involved in the neuroendocrine control of reproduction. Neuroendocrinology 2011; 93:74-89.
44. Cornet A. Current challenges in contraception in adolescents and young women. Curr Opin Obstet Gynecol 2013; 25:S1-10.
45. Castellano JM, Bentsen AH, Sanchez-Garrido MA, Ruiz-Pino F, Romero M, Garcia-Galiano D, et al. Early metabolic programming of puberty onset: impact of changes in postnatal feeding and rearing conditions on the timing of puberty and development of the hypothalamic kisspeptin system. Endocrinology 2011; 152:3396-3408.
46. Roa J, Castellano JM, Navarro VM, Handelsman DJ, Pinilla L, Tena-Sempere M. Kisspeptins and the control of gonadotropin secretion in male and female rodents. Peptides 2009; 30:57-66.
47. Rometo AM, Krajewski SJ, Voytko ML, Rance NE. Hypertrophy and increased kisspeptin gene expression in the hypothalamic infundibular nucleus of postmenopausal women and ovariectomized monkeys. J Clin Endocrinol Metab 2007; 92:2744-2750.
48. Lapatto R, Pallais JC, Zhang D, Chan YM, Mahan A, Cerrato F, et al. Kiss1-/- mice exhibit more variable hypogonadism than Gpr54-/- mice. Endocrinology 2007; 148:4927-4936.
49. Sagheb MM, Azarpira N, Mokhtary M. The effect of ghrelin on Kiss-1 and KissR gene transcription and insulin secretion in rat islets of Langerhans and CRI-D2 cell line. Iran J Basic Med Sci 2017; 20:36-40.
50. Naule L, Robert V, Parmentier C, Martini M, Keller M, Cohen-Solal M, et al. Delayed pubertal onset and prepubertal Kiss1 expression in female mice lacking central oestrogen receptor beta. Hum Mol Genet 2015; 24:7326-7338.
51. Kauffman AS, Gottsch ML, Roa J, Byquist AC, Crown A, Clifton DK, et al. Sexual differentiation of Kiss1 gene expression in the brain of the rat. Endocrinology 2007; 148:1774-1783.
52. Kauffman AS, Navarro VM, Kim J, Clifton DK, Steiner RA. Sex differences in the regulation of Kiss1/NKB neurons in juvenile mice: implications for the timing of puberty. Am J Physiol Endocrinol Metab 2009; 297:E1212-1221.