Chronic Effects of Di(2-ethylhexyl)phthalate on Stereological Parameters of Testis in Adult Wistar Rats

Document Type : Original Article

Authors

1 Department of Biology, Faculty of Sciences, Shahid Chamran University of Ahwaz, Ahwaz,

2 Department of Anatomical Sciences, Faculty of Medicine, Jondishapour Medical Sciences University of Ahwaz, Ahwaz, Iran

Abstract

Objective(s)
In recent years concerns have been raised regarding the incidence of male reproductive disorders from exposure to endocrine disruptors. So, chronic effects of di(2-ethylhexyl)phthalate were studied on histological and stereological structure of testis in adult Wistar rats.
Materials and Methods
Thirty two adult Wistar rats were randomly divided in four equal experiment groups; oil vehicle group and three treated groups which received 10, 100 and 500 mg/kg/day di(2-ethylhexyl)phthalate by gavage for 90 days, respectively. At the end of exposure period the volume of testes was measured by Cavellieri method, testes weight was recorded and then fixed in Bouin’s solution. Following tissue processing, 5 pm sections were stained with haematoxylin-eosin and evaluated with quantitative techniques. Seminiferous tubule diameter, germinal epithelium height, relative and total volumes of seminiferous tubules, tubular lumen and interstitial tissue were estimated.
Results
The results showed that mean weight and volume of testis were decreased significantly (35.2% and 23.9% respectively) in rats treated with 500 mg/kg/day DEHP for 90 days. Seminiferous tubules diameter reduced, 4.4% and 13.4% in 100 and 500 mg/kg/day DEHP-treated groups, respectively. Relative volumes of tubular lumen and interstitial tissue were increased significantly in 100 (P< 0.05) and 500 (P< 0.01) mg/kg/day doses groups. Also, testosterone serum levels were significantly higher (P< 0.05) in rats exposed to 500 mg/kg/day DEHP.
Conclusion
Present study indicated dose-dependent reductions of testicular parameters in adult male rats chronically exposed to di(2-ethylhexyl)phthalate.

Keywords

References

1.Brake A, Krause W. Decreasing quality of semen. BMJ 1992; 305:1498.
2.Suresh C, Sikka RW. Endocrine disruptors and estrogenic effects on male reproductive axis. Asian J Androl 2008; 10:134-145.
3.Kurahashi N, Kondo T, Omura M, Umemura T, Ma M, Kishi R. The effects of subaccute inhalation of Di(2- ethylhexyl) Phthalate on the testes of prepupertal wistar rats. J Occup Health 2005; 47:437-444.
4.Akingbemi BT, Youker RT, Sottas CM, Ge R, Katz E, Klinefelter GR, et al. Modulation of rat Leydig cell steroidogenic function by di(2-ethylhexyl)phthalate. Biol Reprod 2001; 65:1252-1259.
5.Arcadi FA, Costa C, Imperatore C, Marchese A, Rapisardi A, Salemi M, et al. Oral toxicity of di(2- ethylhexyl)phthalate during pregnancy and suckling in the Long-Evans rat. Food Chem_Toxicol 1998; 963-97.
6.Poon R, Lecavaliert P, Mueller R, Valli VE, Procter BG, Chu I. Subchronic oral toxicity of di(n-octyl) phthalate and di(2-ethylhexyl) phthalate in the rat. Food Chem Toxicol 1997; 35:225-239.
7.David RM, Moore MR, Finney DC, Guest D. Chronic toxicity of di(2-ethylhexyl)phthalate in rats. Toxicol Sci 2000; 55:433-443.
8.Lamb JC, Chapin RE, Teague J, Lawton AD, Reel JR. Reproductive effects of four phthalic acid esters in the mouse. Toxicol Appl Pharmacol 1987; 88:55-269.
9.Ge RS, Chen GR, Dong Q, Akingbemi B, Sottas CM, Santos M, et al. Biphasic effects postnatal exposure to di(2-ethylhexyl) phthalate on the timing of puberty in male rats. J Androl 2007; 28: 513-520.
10.Sprando RL. Perfusion of the rat testis through the heart using heparin. In: Russell LD, Ettlin RA, Sinha HAP, Clegg ED, editors. Histological and Histopathological Evaluation of the Testis. Clearwater: Cache River Press; 1990. p. 277-280.
11.Howard CV, Reed MG. Unbiased Stereology. Guilford: Bios Scientific Publishers; 1998.
12.Creasy DM, Foster JR, Foster PMD. The morphological development of di (n-penthyl) phthalate induced testicular atrophy in the rat. J Pathol 1983; 139:309-321. 
13.Gray TJB, Gangoli SD. Aspects of the testicular toxicity of Phthalate esters. Environ Health Piespect 1986; 65:229-235.
14.Oishi S. Prevention of di(2-ethylhexyl)phthalate-induced testicular atrophy in rats by co-administration of the vitamin B12 derivative adenosylcobalamin. Arch Environ Contam Toxicol 1994; 26:497-503.
15.Rhodes C, Orton TC, Pratt IS. Compartive pharmaocokinetics and subacute toxicity of di(2- ethylhexyl)phthalate in rats and marmosets: Extrapolation of effects in rodents to man. Environ Health Perspect 1986; 65:299-308.
16.Parmar D, Srivastava SP, Singh GB, Seth PK. Effect of testosterone on the testicular atrophy_caused by di(2- ethylhexyl) phthalate (DEHP). Toxicol Lett 1987; 36:297-308.
17.Klimisch HJ, Gamer AO, Hellwig J, Kaufmann W, Jackh R. di(2-ethylhexyl)phthalate (DEHP): A short-term repeated inhalation toxicity study including fertility assessment. Food Chem_Toxic 1992; 30: 915-919.
18.Pugh G, Isenberg JS, Kamendulis LM, Ackley DC, Clare LJ, Brown R, et al. Effects of Di (isononyl) Phthalate, di(2-ethylhexyl)phthalate, and Clofibrate in cynomolgous monkeys. Toxicol_Sci 2000; 56:181-188.
19.Sjoberg P, Bondesson U, Gray TJB, Ploen L. Effects of di(2-ethylhexyl)phthalate and five of its metabolites on rat testis in vivo and in vitro. Acta Pharmacol Toxicol 1986; 58:225-233.
20.Zacharewski TR, Meek MD, Clemons JH, Wu ZF, Fielden MR, Matthews JB. Examination of the in vitro and in vivo estrogenic activities of eight commercial phthalate esters. Toxicol Sci 1998; 46:282-293.
21.Gray LE, Kelce WR, Wiese T, Tyl R, Gaido K, Cook J, et al. Endocrine screening methods workshop report: detection of estrogenic and androgenic hormonal and antihormonal activity for chemicals that act via receptor or steroidogenic enzyme mechanisms. Reprod Toxicol 1997; 11: 719-750.
22.Akingbemi BT, Ge R, Klinefelter GR, Zirkin BR, Hardy PM. Phthalate-induced Leydig cell hyperplasia is associated with multiple endocrine disturbances. PNAS 2004; 101: 775-780.
23.Hardy MP, Gelber SJ, Zhou ZF, Penning TM, Ricigliano JW, Ganjam VK, et al. Hormonal control of Leydig cells differentiation. Ann N Y Acad Sci 1991; 637:152-163.
24.Oishi S, Higara K. Effect of phthalic acid esters on gonadal function in male rats. Bull Environ Contam Taxicol 1979; 21:65-67.
25.Ljungvall K, Karlsson P, Hulten F, Madej A, Norrgren L, Einarsson S, et al. Delayed effects on plasma concentration of testosterone and testicular morphology by intramuscular low-dose di(2-ethylhexyl)phthalate or estradiol benzoate in the prepubertal boar. Theriogenology 2005; 64: 1170-1184.
26.Foster PM, Mylchreest E, Gaido KW, Sar M. Effects of phthalate esters on the developing reproductive tract of male rats. Hum Reprod Update 2001; 7:231-235.
27.Shultz VD, Phillips S, Sar M, Foster PM, Gaido KW. Analysis of altered gene expression profiles in fetal rat testis exposed to di (n-butyl) phthalate in utero using cDNA arrays. Taxicol Sci 2001; 64:233-242.
Iranian Journal of Basic Medical Sciences
Volume 13, Issue 4 - Serial Number 4
October 2010
Pages 170-176

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