Gökhan NUR, Sevda YILDIZ ELİŞ, Mümtaz NAZLI

Immunohistochemical distribution and gene expression of transforming growth factor alpha in ovarian tissue of rats treated with capsaicin in puberty*

Aim: In this study, the effects of capsaicin on the immunohistochemical distribution and gene expression of transforming growth factor alpha (TGF-α) in ovarian tissue of rats at the pubertal age were examined. Materials and methods: Rats (50 days old) were divided into experimental, alimentation, and sham groups. The experimental group was injected with 1 mg/kg doses of capsaicin (10% ethanol and 1% Tween in an 80% distilled water mixture) subcutaneously every day for a period of 1 week. The alimentation group received 1 mg/kg doses of capsaicin (10% ethanol and 1% Tween in an 80% distilled water mixture). The sham group was injected with a solution of 10% ethanol and 1% Tween in a mixture of 80% distilled water. Body weights and ovary weights were compared within the groups. Triple staining methods were used for the histological examinations. The tissue localization of TGF-α was analyzed using an immunohistochemical method and gene expression level was determined using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Results: No difference in the structure of the 3 groups was observed histologically. Immunohistochemically, TGF-α immunoreactivity was detected in the primordial follicles, primary follicles, theca cells, and interstitial cells at a medium density in all groups. The effect of capsaicin on TGF-α with mRNA was not found to be significantly different among the 3 groups. Conclusion: The results showed that capsaicin had no effects on ovaries at different developmental stages.

PDF

___

1. Gentry LE, Twardzik DR, Lim GJ, Ranchalis JE, Lee DC. Expression and characterization of transforming growth factor α precursor protein in transfected mammalian cells. Mol Cell Biol 1987; 7: 158591.
2. Marquardt H, Hunkapiller, MW, Hood LE, Todaro, GJ. Rat transforming growth factor type 1: structure and relation to epidermal growth factor. Science 1984; 223:107982.
3. Tapanainen J, Leinonen PJ, Tapanainen P, Yamamoto M, Jaffe RB. Regulation of human granulosa-luteal cell progesterone production and proliferation by gonadotropins and growth factors. Fertil Steril 1987; 48: 57680.
4. Akkoyunlu G, Demir R, Üstünel İ. Distribution patterns of TGF-α, laminin and fibronectin and their relationship with folliculogenesis in rat ovary. Acta Histochem 2003; 105: 295 301.
5. Surh YJ. Anti-tumor promoting potential of selected spice ingredients with antioxidative and anti-inflammatory activities: a short review. Food Chem Toxicol 2002; 40: 10917.
6. Zık B, Erdost H. Histological investigations on the effects of feeding with a diet containing red hot pepper on the uropygial gland of a rooster. Turk J Vet Anim Sci 2002; 26: 122332.
7. Srinivasan MR, Satyanarayana MN. Effect of capsaicin on skeletal muscle lipoprotein lipase in rats fed high fat diet. Indian J Exp Biol 1989; 27: 9102.
8. Hoffmann P, Mazurkiewicz J, Holtmann G, Gerken G, Eysselein VE, Goebell H. Capsaicin-sensitive nerve fibres induce epithelial cell proliferation, inflammatory cell immigration and transforming growth factor-alpha expression in the rat colonic mucosa in vivo. Scand J Gastroenterol 2002; 37: 41422.
9. Abdel Salam OME, Heikal OA, El-Shenawy SM. Effect of capsaicin on bile secretion in the rat. Pharmacology 2005; 73: 1218.
10. Özer A, Zık B, Erdost H, Özfiliz N. Histological investigations on the effects of feeding with a diet containing red hot pepper on the reproductive system organs of the cock. Turk J Vet Anim Sci 2006; 30: 715.
11. Erdost H, Özfiliz N, Özgüden C, Güneş N, Önen S, İlhan T et al. Expression of a capsaicin receptor (VR1) in the testes of mice after an application of capsaicin. Bull Vet Inst Pulawy 2007; 51: 64953.
12. Özgüden Akkoç CG. The immunohistochemical expression of TGF-β in capsaicin injected mice testis at the postnatal development period. PhD, Uludağ University, Bursa, Turkey, 2007.
13. İlhan T. Immunohistochemical expression of ghrelin in mouse testes fed with a diet containing capsaicin on postnatal development. PhD, Uludağ University, Bursa, Turkey, 2009.
14. Zık B, Özgüden Akkoç CG, Tütüncü Ş. The effects of low dose capsaicin on NF-kB and XIAP proteins expression in rat ovary. Ankara Üniv Vet Fak Derg 2010; 57: 2238.
15. Arslan SO. Morphine modulates microvascular leakage dosedependently in the airway of ovalbumin-sensitized rats. Turk J Med Sci 2010; 40: 27986.
16. Zik B, Altunbas K, Tutuncu S, Ozden O, Ozguden Akkoc CG, Peker S et al. Effects of capsaicin on nitric oxide synthase isoforms in prepubertal rat ovary. Biotech Histochem 2012; 87: 21825.
17. Kawada T, Hagihara K, Iwai K. Effects of capsaicin on lipid metabolism in rats fed a high fat diet. J Nutr 1986; 116: 12728.
18. Kempaiah RK, Srinivasan K. Influence of dietary curcumin, capsaicin and garlic on the antioxidant status of red blood cells and the liver in high-fat-fed rats. Ann Nutr Metab 2004; 48: 31420.
19. Moran C, Morales L, Razo RS, Apolonio J, Quiroz U, Chavira R et al. Effects of sensorial denervation induced by capsaicin injection at birth or on day three of life, on puberty, induced ovulation and pregnancy. Life Sci 2003; 73: 211325.
20. Tütüncü Ş. Immunohistochemical expression of VR1 in ovaries of capsaicin treated rats during development. PhD, Uludağ University, Bursa, Turkey, 2009.
21. Shu S, Ju G, Fan L. The glucose oxidase-DAB-nickel method in peroxidase histochemistry of the nervous system. Neurosci Lett 1988; 85: 16971.
22. Luna LG. Manual of histologic staining methods of the Armed Forces Institute of Pathology, 3rd ed. New York: McGraw-Hill; 1968.
23. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 1569.
24. Boillee S, Cadusseau J, Coulpier M, Grannec G, Junier M. Transforming growth factor α: a promoter of motoneuron survival of potential biological relevance. J Neurosci 2001; 21: 707988.
25. Altunışık R, Coşkun R, Bayraktaroğlu S, Yıldırım E. Sosyal Bilimlerde Araştırma Yöntemleri: SPSS Uygulamalı, 3rd ed. Adapazarı, Turkey: Sakarya Kitabevi; 2004.
26. Lobb DK. Expression and actions of transforming growth factors during human follicular development. Fertil Steril 2009; 92: 10804.
27. Ozcakir HT, Ozbilgin MK, Uyar Y, Lacin S, Caglar H. Immunohistochemical detection of transforming growth factor-alpha, epidermal growth factor, and vascular endothelial growth factor expression in hyperstimulated rat ovary. Acta Obstet Gynecol Scand 2005; 84: 88793.
28. Reeka N, Berg FD, Brucker C. Presence of transforming growth factor alpha and epidermal growth factor in human ovarian tissue and follicular fluid. Hum Reprod 1998; 13: 2199205.
29. Singh B, Armstrong DT. Transforming growth factor α gene expression and peptide localization in porcine ovarian follicles. Biol Reprod 1995; 53: 142935.
30. Kudlow JE, Kobrin MS, Purchio AF, Twardzik DR, Hemandez ER, Asa SL, et al. Ovarian transforming growth factor-alpha gene expression: immunohistochemical localization to the theca-interstitial cells. Endocrinology 1987; 121: 15779.
31. Skinner MK, Coffey RJ Jr. Regulation of ovarian cell growth through the local production of transforming growth factoralpha by theca cells. Endocrinology 1988; 123: 26328.
32. Yeh J, Lee GY, Anderson E. Presence of transforming growth factor-alpha messenger ribonucleic acid (mRNA) and absence of epidermal growth factor mRNA in rat ovarian granulosa cells, and effects of these factors on steroidogenesis in vitro. Biol Reprod 1993; 48: 107181.
33. Harada T, Fujikawa T, Yoshida S, Onohara Y, Tanikawa M, Terakawa N. Expression of transforming growth factor alpha (TGF-α) gene in mouse embryonic development. J Assist Reprod Genet 1997; 14: 2629.
34. Pehlivan T, Mansour A, Spaczynski RZ, Duleba AJ. Effects of transforming growth factors-alpha and -beta on proliferation and apoptosis of rat theca-interstitial cells. J Endocrinol 2001; 170: 63945.