Localization of Laminin and Fibronectin in Rat Testes after Diisobutyl Phthalate Exposure: Histopathologic and Immunohistochemical Study

Abstract

In this study the effects of Diisobutyl phthalate (DIBP) from commonly used pytalates were histopathologically investigated in order to find out whether it is one of the causes of increased infertility in recent years. In addition, the localization of laminin and fibronectin was demonstrated immunohistochemically after exposure to DIBP. For this purpose, three study groups from Wistar albino rats were created: experimental group, control group and corn oil applied group. The experimental group were administered by gavage 3 different doses (0.25-0.5-1 ml/kg/day) of DIBP mixed with corn oil every day for 28 days. At the end of the experiment, the testis tissue samples from all of the experimental and control group animals were investigated and evaluated with a light microscope after routine histological and immunohistochemical preparation processes. Reduction in spermatogenic cell population, apoptotic and necrotic germ cells, increase in Sertoli cells, multinucleate giant cells, thickening and dilatation in basal lamina of seminiferous tubules, oedema and shrinkage in interstitial connective tissue were observed in DIBP treatment groups. The decrease in localization of fibronectin and laminin immunoreactivity in seminiferous tubules was observed. The results of this study demonstrated that DIBP causes serious damage to the testicular tissue, especially in terms of spermatogenesis. For this reason, it has come to the conclusion that the material in question may cause infertility in male individuals.

Keywords

• Diisobutyl phthalate,infertility,testes,laminin,fibronectin,histopathology

References

1. 1. Chemical Book, Diisobutyl phthalate Chemical Properties, Uses, Production. https://www.chemicalbook.com/chemicalproductproperty 2017 (accessed 15.06.2017).
2. 2. Environmental protective Agency (EPA). Phtalates-Diisobutylphthalate, Biomonitoring, America’s Children and the Environment, United States, Third Edition, 2017, pp 168-356.
3. 3. Barlow, NJ, Foster, PM. 2003. Pathogenesis of male reproductive tract lesions from gestation through adulthood follwing in utero exposure to di(n-butyl) phytalate. Toxicolology; 31, 397-410.
4. 4. Veeramachaneni, DN, Parks, L. 2000. Perinatal exposure to the phytalates DEHP, BBP and DINP but DEP, DMP or DOTP, alters sexual diffrentiation of the mal erat. Toxicological Sciences; 58, 350-365.
5. 5. Borch, J, Ladefoged, O, Vinggaard, AM. 2004. Steroidogenesis in fetal male rats is reduced by DEHP and DINP, but endocrine effects of DEHP are not modulate by DEHA in fetal, prebubertal and adult male rats. Reproductive Toxicology; 18, 53-61.
6. 6. Parks, LG, Ostby, JS, Lambright, CR, Abbot, BD, Klinefelter, GR, Barlow, NJ, Gray, JLE. 2000. The plasticizer diethylhexyl phytalate induces malmormations by decreasing fetal testosterone synthesis during sexual differentiation in the male rat. Toxicological Sciences, 58, 339-349.
7. 7. Bosman, FT, Stamenkovic, I.2003. Functional structure and composition of the extracellular matrix. Journal Pathology, 200, 423-428.
8. 8. Humphries, MJ. 2000. Integrin structure, Biochemical Society Transactions; 28, 311-339.

9. 9. OECD, Guidelines for Testing of Chemicals No: 407. Repeated Dose 28-day Oral Toxicity Study in Rodents, Organization for Economic Cooperation and Development, 1995, Paris, France.
10. 10. Bancroft, JD, Cook, HC. Manual of Histological Tecniques and Their Diagnostic Application. New York, Churchill Livingstone, 1994, p 457.
11. 11. Cheng, CY, Mruk, DD. 2002. Cell junction dynamics in the testis: Sertoli–germ cell interactions and male contraceptive development. Physiological Reviews; 82, 825-74.
12. 12. Creasy, DM. 2001. Pathogenesis of male reproductive toxicity. Toxicologic Pathology; 29(1), 64-76.
13. 13. Spade, D.J, Hall, S.J, Wilson, S, Boekelheide, K. 2005. Di-n-Butyl Phthalate Induces Multinucleated Germ Cells in the Rat Fetal Testis Through a Nonproliferative Mechanism. Biology of Reproduction; 93(5), 110.
14. 14. Setchell, BP. 1990. Local control of testicular fluids. Reproduction Fertility and Development; 2, 291-309.
15. 15. Cheng, CY, Siu, MKY. 2008. Extracellular matrix and its role in spermatogenesis. Advances in Experimental Medicine and Biology; 636, 74-91.
16. 16. Skinner, MK. 1991. Cell-Cell Interactions in the Testis. Endocrine Reviews; 12, 45-77.
17. 17. El Quali, H, Leheup, BP, Gelly, JL, Grignon, G. 1991. Laminin ultrastructural immunolocalization in rat testis during ontogenesis. Histochemistry; 95, 241-246.
18. 18. Shaller, J, Glander, HJ, Dethloff, J. 1993. Evidence of β1 integrins and fibronectin on spermatogenic cells in human testis. Human Repropduction; 8, 1873-1878.
19. 19. Nacar, E, Elmas, Ç, Erdoğan D. 2014. Localization of β1 integrin and fibronectin on spermatogenic cells in rat testis. Mustafa Kemal Üniversitesi Tıp Dergisi; 5(20), 28-37.
20. 20. Santamaria, L, Martinez-Onsurbe, P, Paniagua, R, Nistal, M. 1990. Laminin, type IV collagen and fibronectin in normal and cryptorchid human testes. An immunohistochemical study. International Journal of Andrology; 13, 135-14.